Chemical compounds

ABSTRACT

The invention relates to a compound which is a thiazole derivative of Formula (I), or a pharmaceutically acceptable salt thereof, wherein R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , Formula (A), Z, L, X, m, n and p are as defined herein. The compounds are useful in the treatment and prevention of bacterial infection. The invention also relates to combinations of the compound of Formula (I) with further active agents.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a Section 371 of international Application No.PCT/EP2018/069827, filed Jul. 20, 2018, which was published in theEnglish language on Jan. 24, 2019 as International Publication No. WO2019/016393 A1, which claims priority under 35 U.S.C. § 119(b) toEuropean Application No. 17305973.4, filed Jul. 21, 2017, and EuropeanApplication No. 18290003.5, filed Jan. 8, 2018, and European ApplicationNo. 18150903.5, filed Jan. 9, 2018.

FIELD OF THE INVENTION

The present invention relates to compounds which are thiazolederivatives. The compounds of the invention find use in the preventionor treatment of bacterial infection.

The invention also provides such compounds per se and pharmaceuticalcompositions comprising such compounds. The compounds of the inventionare useful as inhibitors of metallo-β-lactamase (MBL) enzymes. Thecompounds of the invention can be used in combination therapy, forexample in combination with one or more antibiotic agents and optionallywith one or more inhibitors of serine-β-lactamase (SBL) enzymes. Suchcombination therapy has particular applications in prevention ortreatment of bacterial infection caused by bacteria which are resistantto treatment by antibiotic agents when administered alone, especiallywhen the resistance is attributable to the presence ofmetallo-β-lactamase and/or serine-β-lactamase enzymes and treatment withβ-lactam antibiotics alone may be unsuccessful. In such cases thecombination therapy can rescue the antibacterial activity of theβ-lactam antibiotic.

BACKGROUND

Bacteria in both clinical and non-clinical settings are becomingincreasingly resistant to conventional antibiotics, and this resistanceis becoming a serious clinical and epidemiological problem for humanhealth. For example, it has been shown that single amino acid mutationsin bacterial DNA-dependent RNA-polymerase can reduce the bindingaffinity of this target enzyme for antibiotics, leading to a highfrequency of resistance (FoR). One approach to addressing FoR that hasbeen previously considered is to develop a single agent that inhibitstwo related bacterial enzymes. Examples of such agents includegepotidacin, which inhibits two similar DNA-processing components ofTopoisomerase II and IV enzymes, (GyrA and ParC) and zoliflodacin, whichinhibits two similar ATP-hydrolysing components of Topoisomerase II andIV enzymes (GyrB and ParE). However, this approach is not alwayssuitable for addressing other forms of resistance, for example whenmicrobial resistance to an antibiotic arises through production of abacterial enzyme able to deactivate the antibacterial drug.

In Gram-negative bacteria, resistance to antibiotics (particularlyβ-lactam antibiotics) often arises from the production by the organismof β-lactamases. β-Lactamase enzymes include both metallo-β-lactamases(MBL) and serine-β-lactamases (SBL). Serine β-lactamase enzymes use anactive serine to hydrolyse β-lactam rings in a covalent mechanism whilethe structurally different metallo-β-lactamase enzymes use Zn metalcoordination and a hydroxide ion to hydrolyse the β-lactam ring. In thefield of bacterial β-lactamase enzymes, in particular the Gram-negativearea and more particular the Enterobacteriaceae, the olderserine-β-lactamase enzymes have been supplemented by the morerecently-evolved metallo-β-lactamases. Resistance of gram-negativebacteria to β-lactam antibiotics therefore especially arises from theproduction by the organism of two types of β-lactamases.

As discussed above, in gram-negative bacteria, resistance to antibioticsoften arises from the production by the organism of β-lactamases,especially metallo-β-lactamases (MBL). MBL are resistance determinantsof increasing clinical relevance. In fact, because of their broad range,potent carbapenemase activity and resistance to inhibitors, theseenzymes can confer resistance to almost all β-lactam antibiotics.

MBLs were first detected in the mid-1960s as carried by mobile DNAelements in species with only low pathogenic potential. However, genesencoding MBL spread among major Gram-negative bacteria during the 1990sand this has led to a health crisis arising from the internationaldissemination of carbapenem-resistant Enterobacteriaceae producing theVIM-type and NDM-type metallo-β-lactamases.

Functional features of these Enterobacteriaceae include potentcarbapenemase activity and resistance to clinical β-lactamase inhibitors(clavulanate and sulfones). The activity against β-lactams differsbetween the different metallo-β-lactamases, and substrate specificitymight vary from a narrow range (eg, the CphA metallo-β-lactamase ofAeromonas hydrophila), to an extended range (eg, the VIM-typemetallo-β-lactamases, which can degrade almost all classes of β-lactamsapart from the monobactams).

There are three major structural subclasses of MBL which sharesubstantial internal diversity. Members of the different subclassesdiffer not only in their high degree of sequence diversity, but also inthe structure of their active sites. In enzymes of subclasses B1 and B3,the active site contains two zinc ions; in members of subclass B2, theactive site contains only one zinc ion.

Acquired metallo-β-lactamases have been detected in strains ofEnterobacteriaceae, Pseudomonas aeruginosa, Acinetobacter baumannii, andother Gram-negative bacteria. Among acquired MBL, almost all the enzymesbelong to subclass B1, which indicates an overall higher propensity formembers of this subclass to be captured and spread with mobile geneticelements than for members of subclasses B2 and B3.

As an example, the subclass B1 comprises the IMP-type, the VIM-type, andthe NDM-type enzymes.

The IMP-type enzymes, including IMP-1, were first detected in Japan inthe late 1980s, and have since been reported worldwide inEnterobacteriaceae and in Gram-negative bacteria. The IMP-type enzymeshave broad substrate specificity with a high affinity for cephalosporinsand carbapenems, but they have little activity against Temocillin.

The VIM-type enzymes, including VIM-2, were first discovered in Europein the late 1990s and have since been reported worldwide. VIM-typeenzymes were initially detected in P. aeruginosa and in otherGram-negative bacteria, but have since emerged in Enterobacteriaceae,and have become a major problem in some settings. More than 20 differentVIM allotypes are known, each with a defined geographical distributionexcept for VIM-1 and VIM-2, which share a broader distribution than theIMP-type enzymes. The VIM-type metallo-β-lactamases show even broadersubstrate specificities than the IMP-types, being able to hydrolyse6-α-methoxy-penicillins. Furthermore, the VIM-type enzymes are unique inthe metallo-β-lactamases in that they have a high affinity forcarbapenems.

New Delhi metallo-β-lactamase 1 (NDM-1) is a novel metallo-β-lactamaseidentified initially in a patient hospitalized in New Delhi with aninfection caused by Klebsiella pneumoniae. Subsequently, organisms inthe Enterobacteriaceae family containing this new β-lactamase have beenfound widely distributed throughout India, Pakistan, and Bangladesh andare now occurring in the United Kingdom and in many other countries. TheNew Delhi metallo-β-lactamase 1 (NDM-1) is a polypeptide of 158 aminoacids in length (Accession number AB571289) capable of hydrolyzing awide range of β-lactam antibiotics including penicillins, cephalosporinsand carbapenem antibiotics that are a mainstay for the treatment ofantibiotic-resistant bacterial infections.

Accordingly, there is an urgent need for new antibacterial compounds andcompositions and adjuvant therapies for treating bacterial infection, inparticular bacterial infection caused by bacteria which express MBLenzymes. There is also an urgent need for new compositions for treatingbacterial infection by bacteria which exhibit high resistance,particularly when resistance to antibiotic agents (especially β-lactamantibiotic agents) arises through production by the bacteria of one ormore enzyme able to deactivate the antibacterial drug. The presentinvention aims to address some or all of these issues.

SUMMARY OF THE INVENTION

Previously, the inventors reported in WO 2014/198849 that certainthiazole derivatives are inhibitors of metallo-β-lactamases, includingNDM-1. The inventors have now surprisingly found that compounds ofFormula (I) are potent inhibitors of metallo-β-lactamases, includingNDM-1, and have improved properties compared to the compounds disclosedin WO 2014/198849. The compounds therefore are useful in treating andpreventing bacterial infection, for example by use in combination withβ-lactam antibiotics.

The inventors have also found that the compounds of Formula (I) canadvantageously be used in combination with inhibitors ofserine-β-lactamase enzymes and other antibiotic agents such as β-lactamantibiotics e.g. carbapenem antibiotics. Such combination therapies haveparticular relevance in the prevention or treatment of bacterialinfection caused by bacteria which exhibit a high degree of resistanceto the antibiotic agents when administered alone, especially when thebacterial infection is caused by bacteria which produce β-lactamaseenzymes.

Accordingly, the invention provides a compound which is a thiazolederivative according to Formula (I), or a pharmaceutically acceptablesalt thereof,

wherein

-   -   R¹ is selected from H, R^(1a) and —CH₂OC(O)R^(1a), wherein        R^(1a) is selected from an unsubstituted C₁ to C₄ alkyl group        and phenyl;    -   {circle around (A)} is a cyclic group selected from C₆ to C₁₀        aryl, 5- to 10-membered heteroaryl, and 4- to 10-membered        carbocyclic and heterocyclic groups;    -   each R² is independently selected from:    -   (i) halo or R⁸.    -   (ii) C₁₋₃ alkyl, O(C₁₋₃ alkyl), S(C₁₋₃ alkyl), SO(C₁₋₃ alkyl) or        SO₂(C₁₋₃ alkyl), any of which may optionally be substituted with        1, 2 or 3 halo substituents and/or one R⁸ substituent; and    -   (iii) NR^(a)C(O)R^(c), and NR^(a)C(O)NR^(b)R^(c), wherein each        R^(a) and R^(b) is independently selected from hydrogen and        unsubstituted C₁₋₂ alkyl and each R^(c) is unsubstituted C₁₋₂        alkyl;    -   and        -   each R⁸ is independently selected from CN, OH,            —C(O)NR^(f)R^(g), —NR^(f)R^(g), —NR¹⁰C(NR¹¹)R¹²,            —C(NR¹⁰)NR¹¹R¹², and —NR¹⁰C(NR¹¹)NR¹²R¹³; wherein each of            R^(f) and R^(g) is independently H or unsubstituted C₁₋₂            alkyl;    -   m is 0, 1, 2 or 3    -   R³ is selected from hydrogen and a C₁ to C₃ alkyl group which is        unsubstituted or is substituted with 1, 2 or 3 substituents        selected from halogen, —OR¹⁰, and —NR¹⁰R¹¹;    -   n is 0 or 1    -   Z is a bond or is selected from —NR¹⁰C(O)—, —C(O)NR¹⁰—,        —NR¹⁰C(O)NR¹¹—, —NR¹⁰C(O)O—, —OC(O)NR¹⁰, —NR¹⁰C(O)S—,        —SC(O)NR¹⁰, —NR¹⁰C(NR¹¹)—, —C(NR¹⁰)NR¹¹—, —NR¹⁰C(NR¹¹)NR¹²—,        —NR¹⁰C(N⁺R¹¹R¹²)—, —C(N⁺R¹⁰R¹¹)NR¹²—, —NR¹⁰C(N⁺R¹¹R¹²)NR¹³—,        —NR¹⁰C(NR¹¹)O—, —OC(NR¹⁰)NR¹¹, —NR¹⁰C(N⁺R¹¹R¹²)O—,        —OC(N⁺R¹⁰R¹¹)NR¹²—, —NR¹⁰C(NR¹¹)S—, —SC(NR¹⁰)NR¹¹,        —NR¹⁰C(N⁺R¹¹R¹²)S—, —SC(N⁺R¹⁰R¹¹)NR¹²—, —C(O)NR¹⁵—,        —NR¹⁰C(O)NR¹⁵—, —OC(O)NR¹⁵, —SC(O)NR¹⁵, —C(NR¹⁰)NR¹⁵—,        —NR¹C(NR¹¹)NR¹⁵—, —C(N⁺R¹⁰R¹¹)NR¹⁵—, —NR¹⁰C(N⁺R¹¹R¹²)NR¹⁵—,        —OC(NR¹⁰)NR¹⁵, —OC(N⁺R¹⁰R¹¹)NR¹⁵—, —SC(NR¹⁰)NR¹⁵, and        —SC(N⁺R¹⁰R¹¹)NR¹⁵—;    -   L is a bond or is selected from C₁₋₄ alkylene, C₂₋₄ alkenylene,        C₂₋₄ alkynylene, C₁₋₃ alkylene-(C₃₋₆cycloalkylene)-C₁₋₃        alkylene, C₁₋₄ alkylene-(C₃₋₆cycloalkylene) and        (C₃₋₆cycloalkylene)-C₁₋₄ alkylene, wherein L is unsubstituted or        is substituted with 1 or 2 substituents selected from halogen,        —OR¹⁰, and —NR¹⁰R¹¹; or L is —C(R¹⁰)═N—;    -   X is a bond or, when L is other than a bond or —C(R¹⁰)═N—, X is        a bond or is selected from —NR¹⁰—, —O—, —NR¹⁰C(NR¹¹)—, and        —C(NR¹⁰)—;    -   p is 0 or 1;    -   R⁴ is selected from H, —CN and C₁ to C₃ alkyl which is        unsubstituted or is substituted with 1, 2 or 3 substituents        selected from halogen, —OR¹⁰, —NR¹⁰R¹¹, and —CN;        -   or R⁴ is joined together with R⁵ to form, together with the            atoms to which they are attached, a 5- to 6-membered            heterocyclic group comprising at least one saturated carbon            atom in the ring, said heterocyclic group being            unsubstituted or substituted with 1 or 2 substituents            selected from unsubstituted C₁ to C₂ alkyl, halogen, —OR¹⁰,            —NR¹⁰R¹¹, and —CN;    -   R⁵ is selected from H, —CN and C₁ to C₃ alkyl which is        unsubstituted or is substituted with 1, 2 or 3 substituents        selected from halogen, —OR¹⁰, —NR¹⁰R¹¹, and —CN;        -   or R⁵ is joined together with R⁴ to form, together with the            atoms to which they are attached, a 5- to 6-membered            heterocyclic group comprising at least one saturated carbon            atom in the ring, said heterocyclic group being            unsubstituted or substituted with 1 or 2 substituents            selected from unsubstituted C₁ to C₂ alkyl, halogen, —OR¹⁰,            —NR¹⁰R¹¹, and —CN;        -   or R⁵ is joined together with R⁶ to form, together with the            atoms to which they are attached, a 5- to 6-membered            heterocyclic group comprising at least one saturated carbon            atom in the ring, said heterocyclic group being            unsubstituted or substituted with 1 or 2 substituents            selected from unsubstituted C₁ to C₂ alkyl, halogen, —OR¹⁰,            —NR¹⁰R¹¹, and —CN;    -   R⁶ is selected from H, —CN and C₁ to C₃ alkyl which is        unsubstituted or is substituted with 1, 2 or 3 substituents        selected from halogen, —OR¹⁰, —NR¹⁰R¹¹, and —CN;        -   or R⁶ is joined together with R⁵ to form, together with the            atoms to which they are attached, a 5- to 6-membered            heterocyclic group comprising at least one saturated carbon            atom in the ring, said heterocyclic group being            unsubstituted or substituted with 1 or 2 substituents            selected from unsubstituted C₁ to C₂ alkyl, halogen, —OR¹⁰,            —NR¹⁰R¹¹, and —CN;        -   or R⁶ is joined together with R⁷ if present to form,            together with the atoms to which they are attached, a 5- to            6-membered heterocyclic group comprising at least one            saturated carbon atom in the ring, said heterocyclic group            being unsubstituted or substituted with 1 or 2 substituents            selected from unsubstituted C₁ to C₂ alkyl, halogen, —OR¹⁰,            —NR¹⁰R¹¹, and —CN;    -   R⁷ if present is selected from H, —CN and C₁ to C₃ alkyl which        is unsubstituted or is substituted with 1, 2 or 3 substituents        selected from halogen, —OR¹⁰, —NR¹⁰R¹¹, and —CN;        -   or R⁷ is joined together with R⁶ to form, together with the            atoms to which they are attached, a 5- to 6-membered            heterocyclic group comprising at least one saturated carbon            atom in the ring, said heterocyclic group being            unsubstituted or substituted with 1 or 2 substituents            selected from unsubstituted C₁ to C₂ alkyl, halogen, —OR¹⁰,            —NR¹⁰R¹¹, and —CN;    -   each R¹⁰, R¹¹, R¹², R¹³ and R¹⁴ is independently H or methyl;    -   each R¹⁵ is independently substituted C₁ to C₄ alkyl or        unsubstituted C₂ to C₄ alkyl, wherein when R¹⁵ is a substituted        alkyl group the alkyl group is substituted with 1, 2 or 3        substituents independently selected from halogen, CN, OR¹⁰ and        —NR¹⁰R¹¹.

The invention also provides a compound of Formula (I) wherein

-   -   R¹, {circle around (A)}, R², m, R³, n, L, X, p, R⁴, R⁵, R⁶, R⁷        (if present), R¹⁰, R¹¹, R¹², R¹³ and R¹⁴ are as defined herein;    -   Z is a bond or is selected from —NR¹⁰C(O)—, —C(O)NR¹⁰—,        —NR¹⁰C(O)NR¹¹—, —NR¹⁰C(O)O—, —OC(O)NR¹⁰, —NR¹⁰C(O)S—,        —SC(O)NR¹⁰, —NR¹⁰C(NR¹¹)—, —C(NR¹⁰)NR¹¹—, —NR¹⁰C(NR¹¹)NR¹²—,        —NR¹⁰C(N⁺R¹¹R¹²)—, —C(N⁺R¹⁰R¹¹)NR¹²—, —NR¹⁰C(N⁺R¹¹R¹²)NR¹³—,        —NR¹⁰C(NR¹¹)O—, —OC(NR¹⁰)NR¹, —NR¹⁰C(N⁺R¹¹R¹²)O—,        —OC(N⁺R¹⁰R¹¹)NR¹²—, —NR¹⁰C(NR¹¹)S—, —SC(NR¹⁰)NR¹¹,        —NR¹⁰C(N⁺R¹¹R¹²)S—, and —SC(N⁺R¹⁰R¹¹)NR¹²—; and    -   R¹⁵ is absent.

The present invention also provides a pharmaceutical compositioncomprising a compound as described herein and optionally furthercomprising an antibiotic agent. The pharmaceutical composition typicallycomprises a compound as described herein together with at least onepharmaceutically acceptable carrier or diluent and optionally furthercomprises (i) an antibiotic agent and/or (ii) a serine-β-lactamaseinhibitor. Also provided is a product comprising a compound as describedherein in combination with an antibiotic agent.

The invention also provides a compound as described herein for use inthe treatment or prevention of bacterial infection in a subject in needthereof. Also provided is a method for treating or preventing bacterialinfection in a subject, which method comprises administering to saidsubject an effective amount of a compound as described herein. Furtherprovided is the use of a compound as described herein in the manufactureof a medicament for use in treating or preventing bacterial infection ina subject.

The invention also provides a product comprising a compound as describedherein together with a serine-β-lactamase inhibitor and an antibioticagent. The product may be used in the treatment or prevention ofbacterial infection in a subject in need thereof, particularly when thebacterial infection is caused by bacteria which are resistant totreatment by the antibiotic agent when administered alone, andespecially when the resistance is attributable to the presence ofmetallo-β-lactamase and/or serine-β-lactamase enzymes. In patientssuffering from or susceptible to infection by such bacteria, treatmentwith β-lactam antibiotics alone may be unsuccessful.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 shows the results of in vivo efficacy studies conducted using thecompound of Example 2 described herein in a mouse model. Data show thesuppression of bacterial infection by K. pneumonia NTBC104 in mice bymeropenem alone compared to [meropenem+Example 2]. Meropenem at 30 mg/kgreduced bacterial load slightly whereas meropenem at 30 mg/kg plusExample 2 at 30 mg/kg significantly reduced the bacterial load comparedto meropenem alone, showing a 1.6 Log₁₀ reduction in CFUs.

FIG. 2 shows cumulative MIC-meropenem potentiation using the compoundsof Example 2 and Example 26 in a panel of clinical Enterobacteriacaestrains expressing NDM enzymes (196 isolates). Data show that at an 8μg/mL concentration of either Example 2 or Example 26, meropenem ispotentiated to the extent that just under 90% of strains exhibit ameropenem MIC of 8 ug/mL. By contrast, the same concentration ofmeropenem alone is only capable of stopping the growth of <1% of thestrains and within the parameters of the experiment the cessation ofgrowth of 90% of all strains could not be achieved with meropenem alone.

DETAILED DESCRIPTION OF THE INVENTION Definitions

As used herein, a C₁ to C₄ alkyl group is a linear or branched alkylgroup containing from 1 to 4 carbon atoms. A C₁ to C₄ alkyl group isoften a C₁ to C₃ alkyl group or a C₂ to C₄ alkyl group. Examples of C₁to C₄ alkyl groups include methyl, ethyl, n-propyl, iso-propyl, n-butyl,iso-butyl, sec-butyl, and tert-butyl. A C₁ to C₃ alkyl group istypically a C₁ to C₂ alkyl group. A C₁ to C₂ alkyl group is methyl orethyl, typically methyl. For the avoidance of doubt, where two alkylgroups are present, the alkyl groups may be the same or different.

As used herein, a C₂-C₄ alkenyl group is a linear or branched alkenylgroup containing from 2 to 4 carbon atoms and having one or more, e.g.one or two, typically one double bonds. Typically a C₂-C₄ alkenyl groupis a C₂-C₃ alkenyl group. Examples of C₂-C₄ alkenyl groups includeethenyl, propenyl and butenyl. For the avoidance of doubt, where twoalkenyl groups are present, the alkenyl groups may be the same ordifferent.

As used herein, a C₂-C₄ alkynyl group is a linear or branched alkynylgroup containing from 2 to 4 carbon atoms and having one or more, e.g.one or two, typically one triple bonds. Typically a C₂-C₄ alkynyl groupis a C₂-C₃ alkynyl group. Examples of C₂ to C₄ alkynyl groups includeethynyl, propynyl and butynyl. For the avoidance of doubt, where twoalkynyl groups are present, the alkynyl groups may be the same ordifferent.

As used herein, a C₁ to C₄ alkylene group is an unsubstituted orsubstituted bidentate moiety obtained by removing two hydrogen atomsfrom a C₁ to C₄ alkane. The two hydrogen atoms may be removed from thesame carbon atom or from different carbon atoms. Typically a C₁ to C₄alkylene group is a C₁ to C₃ alkylene group. Examples of C₁ to C₄alkylene groups include methylene, ethylene, n-propylene, iso-propylene,n-butylene, sec-butylene and tert-butylene. A C₁ to C₄ alkylene group istypically a C₁ to C₂ alkylene group. A C₁ to C₂ alkyl group is methyleneor ethylene, typically methylene. For the avoidance of doubt, where twoalkylene groups are present, the alkylene groups may be the same ordifferent.

As used herein, a C₂ to C₄ alkenylene group is an unsubstituted orsubstituted bidentate moiety obtained by removing two hydrogen atomsfrom a C₂ to C₄ alkene. The two hydrogen atoms may be removed from thesame carbon atom or from different carbon atoms. Typically a C₂ to C₄alkenylene group is a C₂ to C₃ alkenylene group. Examples of C₂ to C₄alkenylene groups include ethenylene, n-propenylene, iso-propenylene,n-butenylene, sec-butenylene and tert-butenylene. A C₂ to C₃ alkenylenegroup is typically a C₂ alkenylene, i.e. ethenylene. For the avoidanceof doubt, where two alkenylene groups are present, the alkenylene groupsmay be the same or different.

As used herein, a C₂ to C₄ alkynylene group is an unsubstituted orsubstituted bidentate moiety obtained by removing two hydrogen atomsfrom a C₂ to C₄ alkyne. The two hydrogen atoms may be removed from thesame carbon atom or from different carbon atoms. Typically a C₂ to C₄alkynylene group is a C₂ to C₃ alkynylene group. Examples of C₂ to C₄alkynylene groups include ethynylene, n-propynylene, iso-propynylene,n-butynylene, sec-butynylene and tert-butynylene. A C₂ to C₃ alkynylenegroup is typically a C₂ alkynylene, i.e. ethynylene. For the avoidanceof doubt, where two alkynylene groups are present, the alkynylene groupsmay be the same or different.

An alkyl, alkenyl, alkynyl, alkylene, alkenylene or alkynylene group asused herein may be unsubstituted or substituted. Unless otherwisestated, substituted alkyl, alkenyl or alkynyl groups typically carry oneor more, e.g. 1, 2, 3 or 4, such as one, two or three e.g. one, or two,e.g. one substituent selected from halogen, —CN, —R⁸, —OR¹⁰, and—NR¹⁰R¹¹, wherein R¹⁰ and R¹¹ are as defined herein. The substituents ona substituted alkyl, alkenyl or alkynyl group are typically themselvesunsubstituted unless otherwise stated. Where more than one substituentis present, these may be the same or different.

As used herein, a halogen is typically chlorine, fluorine, bromine oriodine and is preferably chlorine, bromine or fluorine, especiallychorine or fluorine, especially fluorine.

A 3- to 10-membered carbocyclic group is a cyclic hydrocarbon containingfrom 3 to 10 carbon atoms. A carbocyclic group may be saturated orpartially unsaturated, but is typically saturated. A 3- to 10-memberedpartially unsaturated carbocyclic group is a cyclic hydrocarboncontaining from 3 to 10 carbon atoms and containing 1 or 2, e.g. 1double bond. A 3- to 10-membered carbocyclic group is typically a 4- to10-membered carbocyclic group. Often, a 3- to 10-membered carbocyclicgroup is a 3- to 6-membered carbocyclic group, such as a 4- to6-membered or 5- to 6-membered carbocyclic group. A 3- to 10-memberedcarbocyclic group may be a fused bicyclic group, as defined herein. A 3-to 10-membered carbocyclic group may be a saturated 4- to 6-membered,preferably 5- or 6-membered carbocyclic group. Examples of 3- to6-membered saturated carbocyclic groups include cyclopropyl, cyclobutyl,cyclopentyl and cyclohexyl groups.

A 3- to 10-membered heterocyclic group is a cyclic group containing from3 to 10 atoms selected from C, O, N and S in the ring, including atleast one heteroatom, and typically one or two heteroatoms. Theheteroatom or heteroatoms are typically selected from O, N, and S, mosttypically from S and N, especially N. For example, where theheterocyclic group is denoted a nitrogen-containing heterocyclic group,it contains one nitrogen atom and optionally a further heteroatomselected from O, N and S. A heterocyclic group may be saturated orpartially unsaturated. A 3- to 10-membered partially unsaturatedheterocyclic group is a cyclic group containing from 3 to 10 atomsselected from C, O, N and S in the ring and containing 1 or 2, e.g. 1double bond.

A 3- to 10-membered heterocyclic group is typically a 4- to 10-memberedheterocyclic group. Sometimes a 3- to 10-membered heterocyclic group isa 3- to 6-membered heterocyclic group, such as a monocyclic 4- to6-membered heterocyclic group or a monocyclic 5- or 6-memberedheterocyclic group. Alternatively, a 3- to 10-membered heterocyclicgroup may be a 9- or 10-membered fused bicyclic heterocyclic group (i.e.a fused heterobicyclic group).

Examples of 5- and 6-membered saturated heterocyclic groups includepiperazine, piperidine, morpholine, 1,3-oxazinane, pyrrolidine,imidazolidine, and oxazolidine, including quaternised derivativesthereof, as defined herein. Examples of 5- and 6-membered partiallysaturated heterocyclic groups include tetrahydropyrazine,tetrahydropyridine, dihydro-1,4-oxazine, tetrahydropyrimidine,dihydro-1,3-oxazine, dihydropyrrole, dihydroimidazole anddihydrooxazole, including quaternised derivatives thereof, as definedherein.

Examples of 9- and 10-membered fused heterobicyclic groups include9-membered fused heterobicyclic groups such as indoline,2,3-dihydrobenzofuran, 2,3-dihydrobenzo[b]thiophene,2,3-dihydro-1H-benzo[d]imidazole, 2,3-dihydrobenzo[d]oxazole,2,3-dihydrobenzo[d]thiazole, benzo[d][1,3]dioxole,4,5,6,7-tetrahydrothiazolo[5,4-c]pyridine and4,5,6,7-tetrahydrothiazolo[4,5-c]pyridine, including quaternisedderivatives thereof, as defined herein; and 10-membered heterobicyclicgroups such as 1,2,3,4-tetrahydroquinoline,1,2,3,4-tetrahydroisoquinoline, chromane, isochromane, thiochromane,isothiochromane, 1,2,3,4-tetrahydroquinoxaline,1,2,3,4-tetrahydroquinazoline, 1,4-dihydro-2H-benzo[d][1,3]oxazine,3,4-dihydro-2H-benzo[b][1,4]oxazine,3,4-dihydro-2H-benzo[b][1,4]thiazine,1,4-dihydro-2H-benzo[d][1,3]thiazine, 4H-benzo[d][1,3]dioxine and2,3-dihydrobenzo[b][1,4]dioxine, including quaternised derivativesthereof. Often, the fused heterobicyclic group comprises 1, 2 or 3,preferably 1 or 2 nitrogen atoms.

For the avoidance of doubt, references to a heterocyclic group alsoinclude fused polycyclic ring systems, including for instance fusedbicyclic systems in which a heterocyclic group is fused to an arylgroup. When the heterocyclic group is such a fused heterocyclic group,preferred examples are fused ring systems wherein a 5- to 6-memberedheterocyclic group is fused to a phenyl group.

As used herein, a C₃ to C₆ cycloalkylene group (also referred to as aC₃₋₆cycloalkylene group) is an unsubstituted or substituted bidentatemoiety obtained by removing two hydrogen atoms from a saturated C₃ to C₆carbocyclic group as defined herein. The two hydrogen atoms may beremoved from the same carbon atom or from different carbon atoms.Examples of C₃₋₆cycloalkylene groups include cyclopropylene,cyclobutylene, cyclopentylene and cyclohexylene.

As used herein, a C₆ to C₁₀ aryl group is a substituted orunsubstituted, monocyclic or fused polycyclic aromatic group containingfrom 6 to 10 carbon atoms in the ring portion. Examples includemonocyclic groups such as phenyl and fused bicyclic groups such asnaphthyl and indenyl. Phenyl (benzene) is preferred.

As used herein, a 5- to 10-membered heteroaryl group is a substituted orunsubstituted monocyclic or fused polycyclic aromatic group containingfrom 5 to 10 atoms in the ring portion, including at least oneheteroatom, for example 1, 2 or 3 heteroatoms, typically selected fromO, S and N. A heteroaryl group is typically a 5- or 6-memberedheteroaryl group or a 9- or 10-membered heteroaryl group, preferably a5- or 6-membered heteroaryl group. Preferably, the heteroaryl groupcomprises 1, 2 or 3, preferably 1 or 2 nitrogen atoms.

Examples of 5- and 6-membered heteroaryl groups include pyrrole, furan,thiophene, imidazole, oxazole, thiazole, pyridine, pyridazine,pyrimidine, and pyrazine. Examples of 9- and 10-membered heteroarylgroups include 9-membered heteroaryl groups such as indole,benzothiophene, benzofuran, benzoxazole, benzothiazole, benzimidazole,imidazo[1,2-a]pyridine, [1,2,4]triazolo[1,5-a]pyridine andimidazo[1,2-a]pyrazine, including quaternised derivatives thereof; and10-membered heteroaryl groups such as quinoline, isoquinoline,quinazoline, and quinoxaline.

For the avoidance of doubt, references to a heteroaryl group alsoinclude fused polycyclic ring systems, including for instance fusedbicyclic systems in which a heteroaryl group is fused to an aryl group.When the heteroaryl group is such a fused heteroaryl group, preferredexamples are fused ring systems wherein a 5- to 6-membered heteroarylgroup is fused to a phenyl group.

As used herein, a fused bicyclic group is a group comprising two cyclicmoieties sharing a common bond between two atoms.

A carbocyclic, heterocyclic, aryl or heteroaryl group may beunsubstituted or substituted as described herein. For example, acarbocyclic, heterocyclic, aryl or heteroaryl group may be unsubstitutedor substituted with 1, 2 or 3, typically 1 or 2 such as e.g. 1substituent. Suitable substituents include, halogen; —CN; OR¹⁰ and—NR¹⁰R¹¹ (wherein R¹⁰ and R¹¹ are as defined herein) unsubstituted C₁ toC₂ alkyl and R² as depicted in Formula (I) and defined herein. Thesubstituents on a substituted carbocyclic, heterocyclic, aryl orheteroaryl group are typically themselves unsubstituted, unlessotherwise stated.

Compounds of the invention may comprise heterocyclic or heteroarylgroups comprising at least one nitrogen atom. In such compounds, saidnitrogen atom(s) are independently selected from secondary, tertiary andquaternary nitrogen atom(s). A quaternary nitrogen atom is present whenthe compound comprises a quaternised derivative of one or moremonocyclic groups or fused bicyclic groups. As used herein, aquaternised derivative of a moiety such as a cyclic moiety is formed bybonding an additional alkyl group to a nitrogen atom in the moiety suchthat the valency of the said nitrogen atom increases from 3 to 4 and thenitrogen atom is positively charged.

As used herein, a pharmaceutically acceptable salt is a salt with apharmaceutically acceptable acid or base. Pharmaceutically acceptableacids include both inorganic acids such as hydrochloric, sulphuric,phosphoric, diphosphoric, hydrobromic or nitric acid and organic acidssuch as oxalic, citric, fumaric, maleic, malic, ascorbic, succinic,tartaric, benzoic, acetic, methanesulphonic, ethanesulphonic,benzenesulphonic or p-toluenesulphonic acid. Pharmaceutically acceptablebases include alkali metal (e.g. sodium or potassium) and alkali earthmetal (e.g. calcium or magnesium) hydroxides and organic bases such asalkyl amines, aralkyl amines and heterocyclic amines. Hydrochloridesalts and acetate salts are preferred, in particular hydrochloridesalts.

In Formula (I), the stereochemistry is not limited. In particular,compounds of Formula (I) containing one or more chiral centre may beused in enantiomerically or diastereoisomerically pure form, or in theform of a mixture of isomers. Further, for the avoidance of doubt, thecompounds of the invention may be used in any tautomeric form.Typically, the agent or substance described herein contains at least50%, preferably at least 60, 75%, 90% or 95% of a compound according toFormula (I) which is enantiomerically or diasteriomerically pure.Typically, a compound of the invention comprises by weight at least 60%,such as at least 75%, 90%, or 95% of a single enantiomer ordiastereomer. Preferably, the compound is substantially optically pure.

For the avoidance of doubt, the terms ‘thiazole derivative’ and‘thiazolyl derivative’ may be used interchangeably and unless otherwiseindicated refer to compounds of the invention, such as compounds ofFormula (I).

Compounds of the Invention

Typically, in Formula (I), R¹ is selected from H and R^(1a). Morepreferably, R¹ is H. R^(1a) is typically an unsubstituted C₁ to C₄ alkylgroup, such as an unsubstituted C₁ to C₂ alkyl group. More preferably,R^(1a) is methyl or t-butyl.

In Formula (I), {circle around (A)} may preferably be a cyclic groupselected from C₆ to C₁₀ aryl and 5- to 10-membered heteroaryl groups.{circle around (A)} is preferably a cyclic group selected from phenyl,5- to 6-membered heteroaryl, and 5- to 6-membered carbocyclic andheterocyclic groups. {circle around (A)} is more preferably selectedfrom phenyl and 5- to 6-membered heteroaryl groups. Still morepreferably, {circle around (A)} is a phenyl.

When {circle around (A)} is a 5- to 10-membered heteroaryl group, it ispreferably a 5- or 6-membered group. When {circle around (A)} is a 4- to10-membered heterocyclic or carbocyclic group, it is preferably a 5- or6-membered group. When {circle around (A)} is a heterocyclic orheteroaryl group, it preferably contains 1, 2 or 3, preferably 1 or 2heteroatoms selected from O, N and S. When {circle around (A)} is aheterocyclic or heteroaryl group, it is preferably a nitrogen-containinggroup. When {circle around (A)} is a fused heteroaryl or heterocyclicgroup, {circle around (A)} preferably comprises a benzene ring fused toa 5- or 6-membered heterocyclic or heteroaryl group as defined herein.

Preferably, {circle around (A)} is selected from phenyl, cyclohexane,piperidine, pyridazine, pyridine and thiazole. More preferably, {circlearound (A)} is selected from phenyl, pyridazine, pyridine and thiazole.Still more preferably, {circle around (A)} is phenyl.

In Formula (I), each R² is independently selected from:

-   -   (i) halo or R⁸.    -   (ii) C₁₋₃ alkyl, O(C₁₋₃ alkyl), S(C₁₋₃ alkyl), SO(C₁₋₃ alkyl) or        SO₂(C₁₋₃ alkyl), any of which may optionally be substituted with        1, 2 or 3 halo substituents and/or one R⁸ substituent; and    -   (iii) NR^(a)C(O)R^(c), and NR^(a)C(O)NR^(b)R^(c), wherein each        R^(a) and R^(b) is independently selected from hydrogen and        unsubstituted C₁₋₂ alkyl and each R^(c) is unsubstituted C₁₋₂        alkyl;        wherein R⁸ is as defined herein.

When an R² group is according to option (i) above, preferably the groupis a halo group. Fluorine is preferred.

When an R² group is according to option (ii) above, preferably the C₁₋₃alkyl group in the R² moiety is a C₁₋₂ alkyl group, more preferably a C₁alkyl (methyl) group. The R² group is preferably selected from C₁₋₂alkyl, O(C₁₋₂ alkyl), S(C₁₋₂ alkyl) and SO(C₁₋₂ alkyl), more preferablyfrom C₁₋₂ alkyl and O(C₁₋₂ alkyl), each of which may be unsubstituted orsubstituted as described above. When an R² group is according to option(ii) above, the R² group may optionally be substituted with 1, 2 or 3halo substituents and/or one R⁸ substituent; preferably with either 1, 2or 3 halo substituents (of which one or more, preferably all arefluorine) or with one R⁸ substituent. Preferred R² groups according tooption (ii) above include C₁₋₂ alkyl and O(C₁₋₂ alkyl) each of which isunsubstituted or is substituted with 3 fluorine substitutents, such as—CF₃, —OCF₃ and —OCH₃. For the avoidance of doubt, when R² is accordingto option (ii) above and is substituted as described above, the one ormore substituent(s) are each preferably present on the alkyl moiety ofthe R² group.

When an R² group is according to option (iii) above, each R^(a) andR^(b) is independently preferably selected from hydrogen and methyl.Each R^(c) is preferably methyl. More preferably, each R^(a) and R^(b)is independently selected from hydrogen and methyl (preferably hydrogen)and R^(c) is methyl.

Preferably, each R⁸ group is independently selected from CN, OH,—C(O)NR^(f)R^(g), —NR^(f)R^(g), wherein each of R^(f) and R^(g) isindependently H or methyl, preferably hydrogen. More preferably, each R⁸group is independently selected from CN and —C(O)NR^(f)R^(g).

Accordingly, in Formula (I), each R² is preferably independentlyselected from:

-   -   halo or R⁸.    -   C₁₋₂ alkyl, O(C₁₋₂ alkyl), S(C₁₋₂ alkyl), SO(C₁₋₂ alkyl) or        SO₂(C₁₋₂ alkyl), any of which may optionally be substituted with        1, 2 or 3 halo substituents and/or one R⁸ substituent; and    -   NR^(a)C(O)R^(c), and NR^(a)C(O)NR^(b)R^(c), wherein each R^(a)        and R^(b) is independently selected from hydrogen and        unsubstituted C₁₋₂ alkyl and each R^(c) is unsubstituted C₁₋₂        alkyl; wherein each R⁸ is independently selected from CN, OH,        —C(O)NR^(f)R^(g), and —NR^(f)R^(g)—; wherein each of R^(f) and        R^(g) is independently H or unsubstituted C₁₋₂ alkyl.

More preferably, in Formula (I), each R² is independently selected from

-   -   halo, CN, OH, —C(O)NR^(f)R^(g), —NR^(f)R^(g); wherein each of        R^(f) and R^(g) is independently H or methyl; and    -   C₁₋₂ alkyl, O(C₁₋₂ alkyl), S(C₁₋₂ alkyl), SO(C₁₋₂ alkyl) any of        which may optionally be substituted with 1, 2 or 3 halo        substituents and/or one substituent selected from CN and OH.

In Formula (I), m is preferably 0, 1 or 2. More preferably, m is 1 or 2.Sometimes m is 1. Sometimes m is 2.

Therefore, in Formula (I), preferably:

-   -   {circle around (A)} is a cyclic group selected from phenyl, 5-        to 6-membered heteroaryl, and 5- to 6-membered carbocyclic and        heterocyclic groups;    -   each R² is independently selected from:        -   halo or R⁸;        -   C₁₋₂ alkyl, O(C₁₋₂ alkyl), S(C₁₋₂ alkyl), SO(C₁₋₂ alkyl) or            SO₂(C₁₋₂ alkyl), any of which may optionally be substituted            with 1, 2 or 3 halo substituents and/or one R⁸ substituent;            and        -   NR^(a)C(O)R^(c), and NR^(a)C(O)NR^(b)R^(c), wherein each            R^(a) and R^(b) is independently selected from hydrogen and            unsubstituted C₁₋₂ alkyl and each R^(c) is unsubstituted            C₁₋₂ alkyl;    -   wherein each R⁸ is independently selected from CN, OH,        —C(O)NR^(f)R^(g), and —NR^(f)R^(g);    -   wherein each of R^(f) and R^(g) is independently H or        unsubstituted C₁₋₂ alkyl.    -   and    -   m is 0, 1 or 2.

More preferably, in Formula (I):

-   -   {circle around (A)} is selected from phenyl, cyclohexane,        piperidine, pyridazine, pyridine and thiazole;    -   each R² is independently selected from        -   halo, CN, OH, —C(O)NR^(f)R^(g), —NR^(f)R^(g); wherein each            of R^(f) and R^(g) is independently H or methyl; and        -   C₁₋₂ alkyl, O(C₁₋₂ alkyl), S(C₁₋₂ alkyl), SO(C₁₋₂ alkyl) any            of which may optionally be substituted with 1, 2 or 3 halo            substituents and/or one substituent selected from CN and OH;    -   and    -   m is 1 or 2.

Typically, in Formula (I), n is 0.

In Formula (I), if n is 1, R³ is preferably selected from hydrogen andan unsubstituted C₁ to C₃ alkyl group such as methyl or ethyl,preferably methyl. More preferably, R³ if present is hydrogen.

Typically, in Formula (I), Z is a bond or is selected from —NR¹C(O)—,—C(O)NR¹⁰—, —NR¹⁰C(O)NR¹¹—, —NR¹⁰C(O)O—, —OC(O)NR¹⁰, —NR¹⁰C(O)S—,—SC(O)NR¹⁰, —NR¹⁰C(NR¹¹)—, —C(NR¹⁰)NR¹¹—, —NR¹⁰C(NR¹¹)NR¹²—,—NR¹⁰C(NR¹¹)O—, —OC(NR¹⁰)NR¹¹, —NR¹⁰C(NR¹¹)S—, —SC(NR¹⁰)NR¹¹,—C(O)NR¹⁵—, —NR¹⁰C(O)NR¹⁵—, —OC(O)NR¹⁵, —SC(O)NR¹⁵, —C(NR¹⁰)NR¹⁵—,—NR¹⁰C(NR¹¹)NR¹⁵—, —OC(NR¹⁰)NR¹⁵, and —SC(NR¹⁰)NR¹⁵, wherein R¹⁰, R¹¹,R¹² and R are as defined herein. Preferably, Z is a bond or is selectedfrom —NR¹⁰C(O)—, —C(O)NR¹⁰—, —NR¹⁰C(O)NR¹¹—, —NR¹⁰C(O)O—, —OC(O)NR¹⁰,—NR¹⁰C(O)S—, —SC(O)NR¹⁰, —NR¹⁰C(NR¹¹)—, —C(NR¹⁰)NR¹¹—,—NR¹⁰C(NR¹¹)NR¹²—, —NR¹⁰C(NR¹¹)O—, —OC(NR¹⁰)NR¹¹, —NR¹⁰C(NR¹¹)S—, and—SC(NR¹⁰)NR¹¹, wherein R¹⁰, R¹¹ and R¹² are as defined herein. Morepreferably, Z is a bond or is selected from —NR¹⁰C(O)—, —C(O)NR¹⁰—,—NR¹⁰C(O)NR¹¹—, —NR¹⁰C(O)O—, —OC(O)NR¹⁰, —NR¹⁰C(O)S—, —SC(O)NR¹⁰,—NR¹⁰C(NR¹¹)—, —C(NR¹⁰)NR¹¹—, and —NR¹⁰C(NR¹¹)NR²—. Still morepreferably, Z is a bond or is selected from —NR¹⁰C(O)—, —C(O)NR¹⁰—,—NR¹⁰C(O)NR¹¹—, —NR¹⁰C(O)O—, —NR¹⁰C(O)S—, and —NR¹⁰C(NR¹¹)—. Mostpreferably, Z is selected from —NR¹⁰C(O)—, —C(O)NR¹⁰—, and—NR¹⁰C(O)NR¹¹—, preferably —NR¹⁰C(O)—.

Typically, each R¹⁵ is independently substituted C₁ to C₃ alkyl orunsubstituted C₂ to C₃ alkyl, more preferably each R¹⁵ is independentlysubstituted C₁ to C₂ alkyl or unsubstituted C₂ alkyl; still morepreferably each R¹⁵ is independently substituted or unsubstituted C₂alkyl. When R¹⁵ is a substituted alkyl group the alkyl group istypically substituted with 1, 2 or 3, preferably 1 or 2, more preferably1 substituents independently selected from halogen, CN, and OR¹⁰, morepreferably from CN and —OR¹⁰, most preferably from CN and OH.

In Formula (I), L is a bond or is selected from C₁₋₄ alkylene, C₂₋₄alkenylene, C₂₋₄ alkynylene, C₁₋₃ alkylene-(C₃₋₆cycloalkylene)-C₁₋₃alkylene, C₁₋₄ alkylene-(C₃₋₆cycloalkylene) and (C₃₋₆cycloalkylene)-C₁₋₄alkylene, wherein L is unsubstituted or is substituted with 1 or 2substituents selected from halogen, —OR¹⁰, and —NR¹⁰R¹¹; or L is—C(R¹⁰)═N—. Typically, in Formula (I), L is unsubstituted or issubstituted with 1 substituent selected from halogen, —OR¹⁰, and—NR¹⁰R¹¹; most typically L is unsubstituted. When L is other than a bondor —C(R¹⁰)═N— and L is substituted by one or more substituents asdescribed above, the one or more substituents are each preferablypresent on the alkylene, alkenylene or alkynylene group(s) of L. For theavoidance of doubt, when L is a bond L is unsubstituted.

L is preferably a bond or is selected from C₁₋₄ alkylene, C₂₋₄alkenylene and C₂₋₄ alkynylene; or L is —C(R¹⁰)═N—. More preferably, Lis a bond or is selected from C₁₋₃ alkylene and C₂₋₃ alkenylene or is—C(R¹⁰)═N—. Still more preferably, L is selected from C₁₋₃ alkylene andC₂₋₃ alkenylene.

Typically, in Formula (I), X is a bond or, when L is other than a bondor —C(R¹⁰)═N—, X is a bond or is selected from —NR¹⁰— and —O—. Morepreferably, X is a bond.

Preferably, therefore, in Formula (I):

-   -   Z is a bond or is selected from —NR¹⁰C(O)—, —C(O)NR¹⁰—,        —NR¹⁰C(O)NR¹¹—, —NR¹⁰C(O)O—, —OC(O)NR¹⁰, —NR¹⁰C(O)S—,        —SC(O)NR¹⁰, —NR¹⁰C(NR¹¹)—, —C(NR¹⁰)NR¹¹—, and —NR¹⁰C(NR¹¹)NR¹²—;    -   L is a bond or is selected from C₁₋₄ alkylene, C₂₋₄ alkenylene        and C₂₋₄ alkynylene; or L is —C(R¹⁰)═N—;    -   and    -   X is a bond.

More preferably, in Formula (I):

-   -   Z is selected from —NR¹⁰C(O)—, —C(O)NR¹⁰—, and —NR¹⁰C(O)NR¹¹—;    -   L is selected from C₁₋₃ alkylene and C₂₋₃ alkenylene each of        which are preferably unsubstituted;    -   and    -   X is a bond.

In Formula (I), R⁴ is:

-   -   (i) selected from H, —CN and C₁ to C₃ alkyl which is        unsubstituted or is substituted with 1, 2 or 3 substituents        selected from halogen, —OR¹⁰, —NR¹⁰R¹¹, and —CN;    -   or    -   (ii) R⁴ is joined together with R⁵ to form, together with the        atoms to which they are attached, a 5- to 6-membered        heterocyclic group comprising at least one saturated carbon atom        in the ring, said heterocyclic group being unsubstituted or        substituted with 1 or 2 substituents selected from unsubstituted        C₁ to C₂ alkyl, halogen, —OR¹⁰, —NR¹⁰R¹¹, and —CN.

In Formula (I), R⁵ is

-   -   (i) selected from H, —CN and C₁ to C₃ alkyl which is        unsubstituted or is substituted with 1, 2 or 3 substituents        selected from halogen, —OR¹⁰, —NR¹⁰R¹¹, and —CN;    -   or    -   (ii) is joined together with R⁴ to form, together with the atoms        to which they are attached, a 5- to 6-membered heterocyclic        group comprising at least one saturated carbon atom in the ring,        said heterocyclic group being unsubstituted or substituted with        1 or 2 substituents selected from unsubstituted C₁ to C₂ alkyl,        halogen, —OR¹⁰, —NR¹⁰R¹¹, and —CN;    -   or    -   (iii) is joined together with R⁶ to form, together with the        atoms to which they are attached, a 5- to 6-membered        heterocyclic group comprising at least one saturated carbon atom        in the ring, said heterocyclic group being unsubstituted or        substituted with 1 or 2 substituents selected from unsubstituted        C₁ to C₂ alkyl, halogen,        -   —OR¹⁰, —NR¹⁰R¹¹, and —CN;

When R⁴ is C₁ to C₃ alkyl according to option (i) above, it is typicallyunsubstituted or is substituted with 1, 2 or 3 halo substituents or with1 or 2 halo substituents and/or with one substituent selected from—OR¹⁰, —NR¹⁰R¹¹, and —CN. When R⁴ is according to option (i) above, R⁴is preferably H or C₁ to C₂ alkyl which is unsubstituted or issubstituted with 1, 2 or 3 halo substituents or with one —OR¹⁰substituent; more preferably R⁴ is H or methyl, most preferably H.

When R⁵ is C₁ to C₃ alkyl according to option (i) above, it is typicallyunsubstituted or is substituted with 1, 2 or 3 halo substituents or with1 or 2 halo substituents and/or with one substituent selected from—OR¹⁰, —NR¹⁰R¹¹, and —CN. When R⁵ is according to option (i) above, R⁵is preferably selected from H, —CN and C₁ to C₂ alkyl which isunsubstituted or is substituted with 1, 2 or 3 halo substituents or one—NR¹⁰R¹¹ substituent; more preferably, R⁵ is H or methyl, mostpreferably H.

When R⁴ is according to option (ii) above and R⁵ is according to option(ii) above so that R⁴ and R⁵ are joined together to form, together withthe atoms to which they are attached, a 5- to 6-membered heterocyclicgroup comprising at least one saturated carbon atom in the ring, theheterocyclic group is preferably unsubstituted or is substituted with 1substituent selected from unsubstituted C₁ to C₂ alkyl, halogen, and—OR¹⁰; more preferably the heterocyclic group is unsubstituted or issubstituted with 1 substituent selected from methyl and methoxy; mostpreferably the heterocyclic group is unsubstituted. Preferably, when R⁴is according to option (ii) above and R⁵ is according to option (ii)above, R⁴ and R⁵ are joined together to form, together with the atoms towhich they are attached, a 5-membered heterocyclic group, preferably4,5-dihydro-1H-imidazole.

In Formula (I), R⁶ is

-   -   (i) selected from H, —CN and C₁ to C₃ alkyl which is        unsubstituted or is substituted with 1, 2 or 3 substituents        selected from halogen, —OR¹⁰, —NR¹⁰R¹¹, and —CN;    -   or    -   (ii) is joined together with R⁵ to form, together with the atoms        to which they are attached, a 5- to 6-membered heterocyclic        group comprising at least one saturated carbon atom in the ring,        said heterocyclic group being unsubstituted or substituted with        1 or 2 substituents selected from unsubstituted C₁ to C₂ alkyl,        halogen, —OR¹⁰, —NR¹⁰R¹¹, and —CN;    -   or    -   (iii) is joined together with R⁷ if present to form, together        with the atoms to which they are attached, a 5- to 6-membered        heterocyclic group comprising at least one saturated carbon atom        in the ring, said heterocyclic group being unsubstituted or        substituted with 1 or 2 substituents selected from unsubstituted        C₁ to C₂ alkyl, halogen, —OR¹⁰, —NR¹⁰R¹¹, and —CN.

When R⁶ is C₁ to C₃ alkyl according to option (i) above, it is typicallyunsubstituted or is substituted with 1, 2 or 3 halo substituents or with1 or 2 halo substituents and/or with one substituent selected from—OR¹⁰, —NR¹⁰R¹¹, and —CN. When R⁶ is according to option (i) above, R⁶is preferably selected from H, —CN and C₁ to C₂ alkyl which isunsubstituted or is substituted with 1, 2 or 3 halo substituents or one—NR¹⁰R¹¹ substituent; more preferably, R⁶ is H or methyl, mostpreferably H.

When R⁵ is according to option (iii) above and R⁶ is according to option(ii) above so that R⁶ and R⁶ are joined together to form, together withthe atoms to which they are attached, a 5- to 6-membered heterocyclicgroup comprising at least one saturated carbon atom in the ring, theheterocyclic group is preferably unsubstituted or is substituted with 1substituent selected from unsubstituted C₁ to C₂ alkyl, halogen, and—OR¹⁰; more preferably the heterocyclic group is unsubstituted or issubstituted with 1 substituent selected from methyl and methoxy; mostpreferably the heterocyclic group is unsubstituted. Preferably, when R⁵is according to option (iii) above and R⁶ is according to option (ii)above, R⁵ and R⁶ are joined together to form, together with the atoms towhich they are attached, a 6-membered heterocyclic group, preferablymorpholine or piperazine, more preferably morpholine.

In Formula (I), p is 0 or 1.

In Formula (I), R⁷ if present is

-   -   (i) selected from H, —CN and C₁ to C₃ alkyl which is        unsubstituted or is substituted with 1, 2 or 3 substituents        selected from halogen, —OR¹⁰, —NR¹⁰R¹¹, and —CN;    -   or    -   (ii) is joined together with R⁶ to form, together with the atoms        to which they are attached, a 5- to 6-membered heterocyclic        group comprising at least one saturated carbon atom in the ring,        said heterocyclic group being unsubstituted or substituted with        1 or 2 substituents selected from unsubstituted C₁ to C₂ alkyl,        halogen, —OR¹⁰, —NR¹⁰R¹¹, and —CN;

When R⁷ is present and is C₁ to C₃ alkyl according to option (i) above,it is typically unsubstituted or is substituted with 1, 2 or 3 halosubstituents or with 1 or 2 halo substituents and/or with onesubstituent selected from —OR¹⁰, —NR¹⁰R¹¹, and —CN. When R⁷ is presentand is according to option (i) above, R⁷ is preferably selected from H,—CN and C₁ to C₂ alkyl which is unsubstituted or is substituted with 1,2 or 3 halo substituents or one —NR¹⁰R¹¹ substituent; more preferably,R⁷ if present is H or methyl, most preferably H.

When R⁷ is present and R⁶ is according to option (iii) above and R⁷ isaccording to option (ii) above so that R⁶ and R⁷ are joined together toform, together with the atoms to which they are attached, a 5- to6-membered heterocyclic group comprising at least one saturated carbonatom in the ring, the heterocyclic group is preferably unsubstituted oris substituted with 1 substituent selected from unsubstituted C₁ to C₂alkyl, halogen, and —OR¹⁰; more preferably the heterocyclic group isunsubstituted or is substituted with 1 substituent selected from methyland methoxy; most preferably the heterocyclic group is unsubstituted.Preferably, when R⁷ is present and is according to option (ii) above andR⁶ is according to option (iii) above, R⁶ and R⁷ are joined together toform, together with the atoms to which they are attached, a 5-memberedheterocyclic group, preferably imidazolidine.

Preferably, therefore, in Formula (I), p is 1 and R⁷ is H or methyl oris joined together with R⁶ to form, together with the atoms to whichthey are attached, an unsubstituted 5- to 6-membered heterocyclic group.Preferably, R⁴ is H or is joined together with R⁵ to form, together withthe atoms to which they are attached, an unsubstituted 5- to 6-memberedheterocyclic group. More preferably, in Formula (I), R⁵ is selected fromH, —CN and C₁ to C₂ alkyl which is unsubstituted or is substituted with1, 2 or 3 halo substituents and/or one —NR¹⁰R¹¹ substituent and R⁶ is Hor methyl. Most preferably, R⁴, R⁵, R⁶ and R⁷ if present are eachselected from methyl and hydrogen, preferably hydrogen.

For the avoidance of doubt, a heterocyclic group comprising at least onesaturated carbon atom in the ring comprises a —CH₂— group within thering, wherein one or both of the hydrogen atoms of the —CH₂— group maybe substituted as defined herein. Usually, the saturated carbon atom inthe ring is unsubstituted; i.e., the a heterocyclic group comprising atleast one saturated carbon atom in the ring usually comprises a —CH₂—group within the ring. A heterocyclic group comprising at least onesaturated carbon atom in the ring is therefore saturated or partiallysaturated. A heterocyclic group comprising at least one saturated carbonatom in the ring is not aromatic.

In some preferred compounds of Formula (I), therefore,

-   -   R¹ is H;    -   {circle around (A)} is a cyclic group selected from phenyl, 5-        to 6-membered heteroaryl, and 5- to 6-membered carbocyclic and        heterocyclic groups;    -   m is 0, 1 or 2;    -   each R² is independently selected from:        -   halo or R⁸;        -   C₁₋₂ alkyl, O(C₁₋₂ alkyl), S(C₁₋₂ alkyl), SO(C₁₋₂ alkyl) or            SO₂(C₁₋₂ alkyl), any of which may optionally be substituted            with 1, 2 or 3 halo substituents and/or one R⁸ substituent;            and        -   NR^(a)C(O)R^(c), and NR^(a)C(O)NR^(b)R^(c), wherein each            R^(a) and R^(b) is independently selected from hydrogen and            unsubstituted C₁₋₂ alkyl and each R^(c) is unsubstituted            C₁₋₂ alkyl;    -   each R⁸ is independently selected from CN, OH; —C(O)NR^(f)R^(g),        and —NR^(f)R^(g); wherein each of R^(f) and R^(g) is        independently H or unsubstituted C₁₋₂ alkyl;    -   n is 0; or n is 1 and R³ is H    -   Z is selected from —NR¹⁰C(O)—, —C(O)NR¹⁰—, —NR¹⁰C(O)NR¹¹—,        —NR¹⁰C(O)O—, —OC(O)NR¹⁰, —NR¹⁰C(O)S—, —SC(O)NR¹⁰, —NR¹⁰C(NR¹¹)—,        —C(NR¹⁰)NR¹¹—, and —NR¹⁰C(NR¹)NR¹²—;    -   L is a bond or is selected from C₁₋₄ alkylene, C₂₋₄ alkenylene        and C₂₋₄ alkynylene; or L is —C(R¹⁰)═N—;    -   X is a bond;    -   i) p is 0;        -   R⁴ is H and R⁵ is selected from H, —CN and C₁ to C₂ alkyl            which is unsubstituted or is substituted with 1, 2 or 3 halo            substituents and/or one —NR¹⁰R¹¹ substituent; or R⁴ is            joined together with R⁵ to form, together with the atoms to            which they are attached, an unsubstituted 5- to 6-membered            heterocyclic group; and        -   R⁶ is H or methyl;    -   or    -   ii) p is 1; and        -   R⁴ is H; R⁵ is selected from H, —CN and C₁ to C₂ alkyl which            is unsubstituted or is substituted with 1, 2 or 3 halo            substituents and/or one —NR¹⁰R¹¹ substituent; R⁶ is H or            methyl and R⁷ is H or methyl; or R⁴ is joined together with            R⁵ to form, together with the atoms to which they are            attached, an unsubstituted 5- to 6-membered heterocyclic            group; R⁶ is H or methyl and R⁷ is H;

In some even more preferred compounds of Formula (I),

-   -   R¹ is H;    -   {circle around (A)} is selected from phenyl, cyclohexane,        piperidine, pyridazine, pyridine and thiazole;    -   m is 1 or 2;    -   each R² is independently selected from:        -   halo, CN, OH, —C(O)NR^(f)R^(g), —NR^(f)R^(g); wherein each            of R^(f) and R^(g) is independently H or methyl; and        -   C₁₋₂ alkyl, O(C₁₋₂ alkyl), S(C₁₋₂ alkyl), SO(C₁₋₂ alkyl) any            of which may optionally be substituted with 1, 2 or 3            substituents selected from halo, CN, OH;    -   n is 0;    -   Z is selected from —NR¹⁰C(O)—, —C(O)NR¹⁰—, and —NR¹⁰C(O)NR¹¹—;    -   L is selected from C₁₋₃ alkylene and C₂₋₃ alkenylene.    -   X is a bond;    -   p is 0; or p is 1 and R⁷ is H;    -   R⁴ is H;    -   R⁵ is selected from H, —CN and C₁ to C₂ alkyl which is        unsubstituted or is substituted with 1, 2 or 3 halo substituents        and/or one —NR¹⁰R¹¹ substituent H; and    -   R⁶ is H.

Particularly preferred compounds of the invention are

-   5-[[4-[(2-guanidinoacetyl)amino]-3-(trifluoromethoxy)phenyl]sulfonylamino]thiazole-4-carboxylic    acid;-   5-[[3-fluoro-4-[[(2-guanidinoacetyl)amino]methyl]phenyl]sulfonylamino]thiazole-4-carboxylic    acid;-   5-[[3-fluoro-4-(guanidinomethyl)phenyl]sulfonylamino]thiazole-4-carboxylic    acid;-   5-[[3-fluoro-4-(2-guanidinoethylsulfanylcarbonylamino)phenyl]sulfonylamino]thiazole-4-carboxylic    acid;-   5-[[4-[2-[(2-amino-2-imino-ethyl)amino]-2-oxo-ethyl]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylic    acid;-   5-[[3-carbamoyl-4-[(2-guanidinoacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylic    acid;-   5-[[3-cyano-4-[(2-guanidinoacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylic    acid;-   5-[[3-fluoro-4-(2-guanidinoethoxycarbonylamino)phenyl]sulfonylamino]thiazole-4-carboxylic    acid;-   5-[(4-guanidinophenyl)sulfonylamino]thiazole-4-carboxylic acid;-   5-[[4-[2-(2-carbamimidoylhydrazino)-2-oxo-ethyl]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylic    acid;-   5-[[3-chloro-4-[(2-guanidinoacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylic    acid;-   5-[[4-[(2-guanidinoacetyl)amino]-3-methoxy-phenyl]sulfonylamino]thiazole-4-carboxylic    acid;-   5-[[4-[[2-(2-carbamimidoylhydrazino)acetyl]amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylic    acid;-   5-[[4-[[(2E)-2-(carbamimidoylhydrazono)acetyl]amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylic    acid;-   5-[[4-[[2-(4,5-dihydro-1H-imidazol-2-ylamino)acetyl]amino]-3,5-difluoro-phenyl]sulfonylamino]thiazole-4-carboxylic    acid;-   5-[[6-[(2-guanidinoacetyl)amino]pyridazin-3-yl]sulfonylamino]thiazole-4-carboxylic    acid;-   5-[[4-[(2-amino-2-imino-ethyl)carbamoylamino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylic    acid;-   5-[[3,5-difluoro-4-(guanidinocarbamoylamino)phenyl]sulfonylamino]thiazole-4-carboxylic    acid;-   5-[[4-[(3-amino-3-imino-propanoyl)amino]-3,5-difluoro-phenyl]sulfonylamino]thiazole-4-carboxylic    acid;-   5-[[4-[[3-(dimethylamino)-3-imino-propanoyl]amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylic    acid;-   5-[[3-fluoro-4-[(2-guanidinooxyacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylic    acid;-   5-[[3-fluoro-4-[[3-imino-3-(methylamino)propanoyl]amino]phenyl]sulfonylamino]thiazole-4-carboxylic    acid;-   5-[[4-[3-(4,5-dihydro-1H-imidazol-2-yl)propanoylamino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylic    acid;-   5-[[2-[(2-guanidinoacetyl)amino]thiazol-5-yl]sulfonylamino]thiazole-4-carboxylic    acid;-   5-[[4-[[2-[(N-cyanocarbamimidoyl)amino]acetyl]amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylic    acid;-   5-[[3-fluoro-4-(guanidinocarbamoylamino)phenyl]sulfonylamino]thiazole-4-carboxylic    acid;-   5-[[3-fluoro-4-[[2-(morpholine-4-carboximidoylamino)acetyl]amino]phenyl]sulfonylamino]thiazole-4-carboxylic    acid;-   5-[[4-[(3-amino-3-imino-2-methyl-propanoyl)amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylic    acid;-   5-[[4-[[2-(4,5-dihydro-1H-imidazol-2-yl)acetyl]amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylic    acid;-   5-[[4-(carbamimidoylcarbamoylamino)-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylic    acid;-   5-[[4-[[(2R)-2-guanidinopropanoyl]amino]phenyl]sulfonylamino]thiazole-4-carboxylic    acid;-   5-[[3,5-difluoro-4-[(2-guanidinoacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylic    acid;-   5-[[4-[(4-amino-4-imino-butanoyl)amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylic    acid;-   5-[[4-[[2-(4,5-dihydro-1H-imidazol-2-ylamino)acetyl]amino]-2,5-difluoro-phenyl]sulfonylamino]thiazole-4-carboxylic    acid;-   5-[[2,5-difluoro-4-[(2-guanidinoacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylic    acid;-   5-[[3-fluoro-4-[[2-[(N-methylcarbamimidoyl)amino]acetyl]amino]phenyl]sulfonylamino]thiazole-4-carboxylic    acid;-   5-[[3-fluoro-4-[[2-(2-iminoimidazolidin-1-yl)acetyl]amino]phenyl]sulfonylamino]thiazole-4-carboxylic    acid;-   5-[[4-[[2-[carbamimidoyl(methyl)amino]acetyl]amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylic    acid;-   5-[[4-[[2-[[N-(2-aminoethyl)carbamimidoyl]amino]acetyl]amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylic    acid;-   5-[[5-fluoro-6-[(2-guanidinoacetyl)amino]-3-pyridyl]sulfonylamino]thiazole-4-carboxylic    acid;-   5-[[3-fluoro-4-(3-guanidinopropanoylamino)phenyl]sulfonylamino]thiazole-4-carboxylic    acid;-   5-[[4-[(3-amino-3-imino-propanoyl)amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylic    acid;-   5-[[3,5-difluoro-4-(guanidinocarbamoylamino)phenyl]sulfonylamino]thiazole-4-carboxylic    acid;-   5-[[3-fluoro-4-[(2-guanidinoacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylic    acid; and-   5-[[4-[(2-guanidinoacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylic    acid; and pharmaceutically acceptable salts thereof.

Synthesis

The compounds of the invention can be prepared by any suitable method.Detailed general synthetic routes for representative compounds of theinvention are set out below and in the Examples.

In summary, compounds of the invention can typically be prepared in areaction according to the following scheme:

Starting material SM is readily available and can, for example, beprepared using the methods described in WO 2014/198849. The disclosureof WO 2014/198849 regarding the formation of compound SM and itsanalogues is incorporated by reference. Reaction of SM with a sulphonylchloride derivative of {circle around (A)} (reaction step 1) yields athiazole sulphonamide derivative of {circle around (A)} (A). Reaction ofA with a W—Z-L-X-Pro moiety (B) yields intermediate C. In the abovescheme, Q and W are complementary reactive groups which react togetherto couple A to B to yield compound C. For example, Q can be bromine and—Z′—W can be —C(O)NH₂ so that Q and W react together via Buchwaldchemistry (this is particularly suited to when n is 0). Alternatively, Qcan be —NH₂ and —Z′—W can be —C(O)OH so that Q and W react together in astandard peptide coupling reaction using reagents such as HATU. Othermethods of coupling compounds are well known to those skilled in theart. In compounds B and C the moiety —NR⁷—Pro represents a protectedamine moiety which can be deprotected to yield the amine via standardmethods such as acid-catalysed deprotection (compounds D). Suitableamine protecting groups are well known to those skilled in the art andinclude Boc (tert-butoxycarbonyl) protecting groups The amine can thenbe reacted to form a guanidine group as in compound E by reaction withknown guanidinylating agents such as 1H-pyrazole-1-carboximidamide. Incompounds of the invention wherein p is zero such that an amidine ratherthan a guanidine group is present, the synthesis shown above can bemodified so that compound B comprises a protected amidine group ratherthan protected amine NR⁷-Pro. Suitable amidine protecting groups arewell known to those skilled in the art and include Boc(tert-butoxycarbonyl) protecting groups. In these cases, reaction of Aand B yields a compound C′ which when deprotected yields the desiredamidine product E′. Detailed synthetic routes to exemplary compounds ofthe invention are set out below.

Therapeutic Efficacy

The compounds of the present invention are therapeutically useful. Thepresent invention therefore provides compounds as described herein, foruse in medicine. The present invention provides compounds as describedherein, for use in treating the human or animal body. For the avoidanceof doubt, the compound of the invention may be administered in the formof a solvate.

Also provided is a pharmaceutical composition comprising a compound ofthe invention together with a pharmaceutically acceptable carrier ordiluent and optionally further comprising an antibiotic agent.Typically, the composition contains up to 85 wt % of a compound of theinvention. More typically, it contains up to 50 wt % of a compound ofthe invention. Preferred pharmaceutical compositions are sterile andpyrogen free. Further, when the pharmaceutical compositions provided bythe invention contain a compound of the invention which is opticallyactive, the compound of the invention is typically a substantially pureoptical isomer.

The composition of the invention may be provided as a kit comprisinginstructions to enable the kit to be used in the methods describedherein or details regarding which subjects the method may be used for.

As explained above, the compounds of the invention are useful intreating or preventing bacterial infection. In particular, they areinhibitors of metallo-β-lactamase (MBL) enzymes and are therefore usefulfor removing or reducing resistance of Gram-negative bacteria toantibiotics.

The compounds of the invention may be used as standalone therapeuticagents. For example, the compounds of the invention may be used asstandalone adjuncts in antibacterial therapy, for example inchemotherapy regimes. Alternatively, they may be used in combinationwith antibiotic agents to enhance the action of the antibiotic agent.The compounds of the invention may find particular use in treating orpreventing bacterial infection caused by bacteria which are resistant totreatment with antibiotic agents when administered alone, particularlywhere the resistance is caused by presence of metallo-β-lactamase and/orserine-β-lactamase enzymes. Treatment or prevention of such infectionwith β-lactam antibiotics alone may be unsuccessful.

The present invention therefore also provides a product comprising (i) acompound of the invention as described herein and (ii) an antibioticagent. The compound of the invention and the antibiotic agent may beprovided in a single formulation, or they may be separately formulated.Where separately formulated, the two agents may be administeredsimultaneously or separately. They may be provided in the form of a kit,optionally together with instructions for their administration. Theproducts may also be referred to herein as combinations orpharmaceutical combinations.

Where formulated together, the two active agents may be provided as apharmaceutical composition comprising (i) a compound of the invention asdescribed herein and (ii) a further antibacterial compound; and (iii) apharmaceutically acceptable carrier or diluent.

Preferably, the antibiotic agent is a β-lactam antibiotic. Morepreferably, the antibiotic agent is a β-lactam antibiotic is selectedfrom carbapenems, penicillins, cephalosporins and penems. Examples ofcarbapenem antibiotics include Imipenem, Meropenem, Ertapenem, Doripenemand Biapenem. Examples of penicillins include Amoxicillin, Ampicillin,Ticarcillin, Piperacillin and Cloxacillin. Examples of cephalosporinsinclude Cefazolin, Ceftriaxone, Ceftazidine and Ceftobiprole. Examplesof penems include Faropenem. Other antibiotic agents include tobramycin,neomycin, streptomycin, gentamycin, tazobactam, rifampicin,ciprofloxacin, amikacin, colistin, aztreonam and levofloxacin.Preferably, the β-lactam antibiotic is a carbapenem antibiotic, morepreferably imipenem or meropenem, most preferably meropenem.

The products of the invention may further comprise a serine-β-lactamase(SBL) inhibitor. Thus, the invention also provides a product comprising(i) a compound of the invention; (ii) a serine-β-lactamase (SBL)inhibitor; and (iii) an antibiotic agent. These products are referred toherein as “triple combinations”. The triple combinations comprise theabove three active agents (i) to (iii) but may also comprise furtheractive agents if desired.

In the triple combination of the invention, the compound of theinvention, the SBL inhibitor and the antibiotic agent may each beprovided in a single formulation, or they may be separately formulated.Alternatively, two of the components may be provided in a singleformulation and the remaining component may be provided separately. Inother words, the compound of the invention may be formulated with theSBL inhibitor and the antibiotic agent; or the compound of the inventionmay be formulated with the SBL inhibitor whilst the antibiotic agent isprovided separately; or the compound of the invention may be formulatedwith the antibiotic agent whilst the SBL inhibitor is providedseparately; or the SBL inhibitor may be formulated with the antibioticagent whilst the compound of the invention is provided separately; orthe compound of the invention, the SBL inhibitor and the antibioticagent may each be formulated separately. Where separately formulated,the components of the triple combination may be administeredsimultaneously or separately. They may be provided in the form of a kit,optionally together with instructions for their administration.

Where two or more active agents are formulated together, the two or moreactive agents may be provided as a pharmaceutical composition comprising(i) a compound of the invention as described herein; (ii) apharmaceutically acceptable carrier or diluent; and one or both of (iii)an antibiotic agent; and (iv) a serine-β-lactamase (SBL) inhibitor.

In the triple combination of the invention, the SBL inhibitor is acompound of Formula (II) or a pharmaceutically acceptable salt thereof,

wherein

-   -   G is selected from —CN and —C(O)NR^(j)R^(k);    -   R^(k) is selected from —W and -Q-W; wherein W is selected from        5- to 6-membered heterocyclyl, R^(j) and —N(R^(j))₂; and Q is        selected from —NR^(j)C(O)—, —C(O)—NR^(j)—, C₁₋₃ alkylene,        —O—C₁₋₃ alkylene and —N(R^(j))—C₁₋₃ alkylene;    -   each R^(j) is selected from H and unsubstituted C₁₋₃ alkyl,        preferably H.

In Formula (II), when W is a 5- to 6-membered heterocyclyl, W ispreferably a 6-membered heterocycle containing a nitrogen atom; morepreferably W is piperidinyl. Preferably, in Formula (II), W is selectedfrom 5- to 6-membered heterocyclyl and —N(R^(j))₂, more preferably W isselected from piperidinyl and NH₂. In formula (II), Q is preferablyselected from —NR^(j)C(O)— and —O—C₁₋₃ alkylene. Preferably, in Formula(II), each R^(j) is H. Thus, preferred definitions G in formula (II) are—CN and —C(O)NHR^(k), wherein R^(k) is selected from —W and -Q-W;wherein W is selected from 5- to 6-membered heterocyclyl, preferablypyridinyl, and —NH₂; and Q is selected from —NHC(O)— and —O—C₁₋₃alkylene.

More preferably, in the pharmaceutical combination of the invention, theSBL inhibitor is selected from WCK4234, avibactam, relebactam,zidebactam and nacubactam, or pharmaceutically acceptable salts thereof.The structures of WCK4234, avibactam, relebactam, zidebactam andnacubactam are shown below. Such SBL inhibitors are commerciallyavailable and/or can be synthesized according to published protocolsavailable to those skilled in the art. For example, WCK4234 and itssynthesis is described in WO 2013/038330 and WO 2015/114595. Avibactamand its synthesis is described in Ball, M. et al, Org. Process Res.Dev., 2016, 20 (10), pp 1799-1805; and US 2012/323010. Relebactam andits synthesis is described in WO 2009/091856. Zidebactam and itssynthesis is described in WO 2015/110885. Nacubactam and its synthesisis described in WO 2014/091268 and US 2016/272641.

More preferably, in the pharmaceutical combination of the invention, theSBL inhibitor is WCK4234 or a pharmaceutically acceptable salt thereof.Still more preferably, the SBL inhibitor is WCK4234 or the sodium saltthereof. A process for the preparation of the sodium salt of WCK4234 isdescribed in WO 2015/114595.

In the triple combination of the invention, the antibiotic agent may beany antibiotic agent disclosed herein. Preferably, in the pharmaceuticalcombination of the invention, the antibiotic agent is a β-lactamantibiotic. Preferably, the β-lactam antibiotic is selected fromcarbapenems, penicillins, cephalosporins and penems, more preferably theβ-lactam antibiotic is a carbapenem antibiotic, preferably imipenem ormeropenem, most preferably meropenem.

Most preferably, therefore, the pharmaceutical combination of theinvention comprises (i) a compound of the invention; (ii) an SBLinhibitor selected from WCK4234, avibactam, relebactam, zidebactam andnacubactam, and pharmaceutically acceptable salts thereof, preferablyWCK4234 or a pharmaceutically acceptable salt thereof; and (iii) acarbapenem antibiotic, preferably meropenem.

The compounds of the invention are also useful in treating or preventingbacterial infection. The present invention therefore provides a compoundof the invention for use in medicine. The invention also provides theuse of a compound of the invention in the manufacture of a medicament.The invention also provides compositions and products comprising thecompounds of the invention, as described here. Such compositions andproducts are also useful in treating or preventing bacterial infection.The present invention therefore provides a composition or product asdefined herein for use in medicine. The invention also provides the useof a composition or product of the invention in the manufacture of amedicament.

As explained above, the compounds, compositions and products of theinvention are useful in treating or preventing bacterial infection. Theinvention therefore also provides a method of treating or preventingbacterial infection in a subject, which method comprises administeringto said subject an effective amount of a compound, composition orproduct as described herein. Further provided is a compound, compositionor product of the invention as described herein for the manufacture of amedicament for use in treating or preventing bacterial infection; thecompound of the invention is often used in combination with anantibiotic agent.

As further explained above, the compounds of the invention are useful incombination with a further antibacterial compound. The inventiontherefore provides a compound of the invention for use in treating orpreventing bacterial infection, wherein such use comprisesco-administering the compound of the invention with a furtherantibacterial compound. The invention also provides the use of acompound of the invention in the manufacture of a medicament fortreating or preventing bacterial infection by co-administration of thecompound of the invention with a further antibacterial compound. Theinvention also provides a method for treating or preventing bacterialinfection by co-administering the compound of the invention and afurther antibacterial compound to a subject in need thereof. The furtherantibacterial compound is preferably an antibacterial compound asdescribed herein; more preferably a β-lactam antibiotic as describedherein.

The compounds of the invention are also useful in combination with aserine-β-lactamase (SBL) inhibitor and an antibiotic agent, i.e. as a“triple combination”. The invention therefore provides a compound of theinvention for use in treating or preventing bacterial infection, whereinsuch use comprises co-administering (i) the compound of the inventionwith (ii) a serine-β-lactamase (SBL) inhibitor and (iii) an antibioticagent. Also provided is an antibiotic agent for use in treating orpreventing bacterial infection by co-administration with a compound ofthe invention and optionally an SBL inhibitor. Also provided is an SBLinhibitor for use in treating or preventing bacterial infection byco-administration with a compound of the invention and optionally anantibiotic agent. The invention also provides the use of a compound ofthe invention in the manufacture of a medicament for treating orpreventing bacterial infection by co-administration of (i) the compoundof the invention with (ii) a serine-β-lactamase (SBL) inhibitor and(iii) an antibiotic agent. Also provided is the use of an antibioticagent in the manufacture of a medicament for use in treating orpreventing bacterial infection by co-administration with a compound ofthe invention and optionally an SBL inhibitor. Also provided is the useof an SBL inhibitor in the manufacture of a medicament for use intreating or preventing bacterial infection by co-administration with acompound of the invention and optionally an antibiotic agent. Theinvention also provides a method for treating or preventing bacterialinfection by co-administering (i) the compound of the invention; and(ii) a serine-β-lactamase (SBL) inhibitor and/or (iii) an antibioticagent, to a subject in need thereof. The serine-β-lactamase (SBL)inhibitor is preferably a serine-β-lactamase (SBL) inhibitor describedherein. The antibiotic agent is preferably an antibacterial compound asdescribed herein; more preferably a β-lactam antibiotic as describedherein.

In one aspect, the subject is a mammal, in particular a human. However,it may be non-human. Preferred non-human animals include, but are notlimited to, primates, such as marmosets or monkeys, commercially farmedanimals, such as horses, cows, sheep or pigs, and pets, such as dogs,cats, mice, rats, guinea pigs, ferrets, gerbils or hamsters. The subjectcan be any animal that is capable of being infected by a bacterium.

The compounds, compositions and combinations described herein are usefulin the treatment of bacterial infection which occurs after a relapsefollowing an antibiotic treatment. The compounds, compositions andcombinations can therefore be used in the treatment of a patient who haspreviously received antibiotic treatment for the (same episode of)bacterial infection.

The bacterium causing the infection may be any bacterium expressing ametallo-β-lactamase enzyme or an analogue thereof. Typically thebacterium causing the infection expresses a MBL enzyme. The bacterium istypically Gram-negative. The bacterium may in particular be a pathogenicbacterium. Typically, the bacterial infection to be treated using thecompounds of the invention is resistant to treatment with a conventionalantibiotic when the conventional antibiotic is used alone.

The Gram-negative bacteria of which antibiotic resistance can be removedusing the compounds of general formula (I) are bacteria which producemetallo-β-lactamases, which may be metallo-β-lactamases of subclassesB1, B2 or B3, for example IMP-type (including IMP-1), VIM-type(including VIM-1 and VIM-2) and NDM-type (including NDM-1) enzymes.Typically, the Gram-negative bacteria express NDM-type MBL enzymes,VIM-type MBL enzymes and/or IMP-type MBL enzymes; more typically thebacteria express NDM-type MBL enzymes and/or VIM-type MBL enzymes; mosttypically the bacteria express NDM-type MBL enzymes. The Gram-negativebacteria may express one or more of the following enzymes: ACT-TYPE,CMY-4, CTX-M-3, CTX-M-15, IMP-1, IMP-28, KPC-2, NDM-1, OXA-48, OXA-181,SHV-OSBL, SHV-11, SHV-12, TEM-OSBL, TEM-1, VIM-1, and/or VIM-19.

The bacterial infection may be caused by bacteria from the familiesEnterobacteriaceae, Pseudomonadaceae and/or Moraxellaceae, moretypically the bacterial infection is caused by bacteria from thefamilies Enterobacteriaceae and/or Pseudomonadaceae, and most typicallythe bacterial infection is caused by bacteria from the familyEnterobacteriaceae. The bacterial infection may be caused by Pseudomonas(e.g. Pseudomonas aeruginosa, Pseudomonas oryzihabitans, or Pseudomonasplecoglossicida), Klebsiella, Escherichia, Acinetobacter orBurkholderia. For example, the bacterial infection may be caused byKlebsiella pneumonia, Escherichia coli, Pseudomonas aeruginosa,Burkholderia cepacia or Acinetobacter baumannii. The bacterial infectionmay be caused by Escherichia coli, Klebsiella pneumonia, or Klebsiellaoxytoca. The bacterium may be an opportunistic pathogen.

The compounds, compositions and products of the invention are useful inthe prevention or treatment of infection by the following strains:

NTBC020 (E. coli strain expressing NDM-1, TEM-1 and CTX-M-15); NTBC035-2(K. pneumoniae strain expressing NDM-1, CMY-4 and SHV-11); NTBC104-1 (K.pneumoniae strain expressing NDM-1 and SHV-11); NTBC123 (K. pneumoniaestrain expressing NDM-1); NTBC062 (K. pneumoniae strain expressing IMP-1and TEM-1); NTBC024 (K. pneumoniae strain expressing VIM-19, TEM-1 andCTX-M-3); NTBC042 (E. coli strain expressing VIM-1, TEM-1, CTX-M-15,SHV-12); NTBC055 (E. Coli strain expressing VIM-1); and NTBC039 (K.oxytoca strain expressing IMP-28).

The compounds, compositions and products of the invention may also beuseful in the prevention or treatment of infection by the followingstrains. The triple combination is particularly useful in the preventionor treatment of infection by these strains:

NTBC019 (K. pneumonia strain expressing NDM-1, CTX-M-15 and OXA-181);NTBC185 (K. pneumonia strain expressing SHV-OSBL, TEM-OSBL, NDM-1 andOXA-48); NTBC186 (K. pneumonia strain expressing ACT-TYPE, VIM-1 andOXA-48); NTBC187 (K. pneumonia strain expressing SHV-OSBL, NDM-1 andOXA-48); and NTBC188 (K. pneumonia strain expressing NDM-1 and KPC-2).

The compound, composition or combination of the invention may be used totreat or prevent infections and conditions caused by any one or acombination of the above-mentioned bacteria. In particular, thecompound, composition or combination of the invention may be used in thetreatment or prevention of pneumonia. The compound, composition orcombination may also be used in the treatment of septic shock, urinarytract infection, and infections of the gastrointestinal tract, skin orsoft tissue.

The compound, composition or combination of the invention may be used totreat patients with Carbapenem Resistant Enterobacteriaceae (CRE). CREcan be found in the community or in hospitals and other institutionswhich are commonly associated with long term patients and those that areundergoing significant medical interventions such as are commonly caredfor in Intensive Care Units (ICUs).

A compound, composition or combination of the invention can beadministered to the subject in order to prevent the onset orreoccurrence of one or more symptoms of the bacterial infection. This isprophylaxis. In this embodiment, the subject can be asymptomatic. Thesubject is typically one that has been exposed to the bacterium. Aprophylactically effective amount of the agent or formulation isadministered to such a subject. A prophylactically effective amount isan amount which prevents the onset of one or more symptoms of thebacterial infection.

A compound, composition or combination of the invention can beadministered to the subject in order to treat one or more symptoms ofthe bacterial infection. In this embodiment, the subject is typicallysymptomatic. A therapeutically effective amount of the agent orformulation is administered to such a subject. A therapeuticallyeffective amount is an amount effective to ameliorate one or moresymptoms of the disorder.

The compound, composition or combination of the invention may beadministered in a variety of dosage forms. Thus, it can be administeredorally, for example as tablets, troches, lozenges, aqueous or oilysuspensions, dispersible powders or granules. Formulation composition ofthe invention may also be administered parenterally, whethersubcutaneously, intravenously, intramuscularly, intrasternally,transdermally or by infusion techniques. The compound, composition orcombination may also be administered as a suppository. Preferably, thecompound, composition or combination may be administered via inhaled(aerosolised) or intravenous administration, most preferably by inhaled(aerosolised) administration.

The compound, composition or combination of the invention is typicallyformulated for administration with a pharmaceutically acceptable carrieror diluent. For example, solid oral forms may contain, together with theactive compound, diluents, e.g. lactose, dextrose, saccharose,cellulose, corn starch or potato starch; lubricants, e.g. silica, talc,stearic acid, magnesium or calcium stearate, and/or polyethyleneglycols; binding agents; e.g. starches, arabic gums, gelatin,methylcellulose, carboxymethylcellulose or polyvinyl pyrrolidone;disaggregating agents, e.g. starch, alginic acid, alginates or sodiumstarch glycolate; effervescing mixtures; dyestuffs; sweeteners; wettingagents, such as lecithin, polysorbates, laurylsulphates; and, ingeneral, non toxic and pharmacologically inactive substances used inpharmaceutical formulations. Such pharmaceutical preparations may bemanufactured in known manner, for example, by means of mixing,granulating, tableting, sugar coating, or film coating processes.

The compound, composition or combination of the invention may beformulated for inhaled (aerosolised) administration as a solution orsuspension. The compound, composition or combination of the inventionmay be administered by a metered dose inhaler (MDI) or a nebulizer suchas an electronic or jet nebulizer. Alternatively, the compound,composition or combination of the invention may be formulated forinhaled administration as a powdered drug, such formulations may beadministered from a dry powder inhaler (DPI). When formulated forinhaled administration, the compound, composition or combination of theinvention may be delivered in the form of particles which have a massmedian aerodynamic diameter (MMAD) of from 1 to 100 μm, preferably from1 to 50 μm, more preferably from 1 to 20 μm such as from 3 to 10 μm,e.g. from 4 to 6 μm. When the compound, composition or combination ofthe invention is delivered as a nebulized aerosol, the reference toparticle diameters defines the MMAD of the droplets of the aerosol. TheMMAD can be measured by any suitable technique such as laserdiffraction.

Liquid dispersions for oral administration may be syrups, emulsions andsuspensions. The syrups may contain as carriers, for example, saccharoseor saccharose with glycerine and/or mannitol and/or sorbitol.

Suspensions and emulsions may contain as carrier, for example a naturalgum, agar, sodium alginate, pectin, methylcellulose,carboxymethylcellulose, or polyvinyl alcohol. The suspension orsolutions for intramuscular injections or inhalation may contain,together with the active compound, a pharmaceutically acceptablecarrier, e.g. sterile water, olive oil, ethyl oleate, glycols, e.g.propylene glycol, and if desired, a suitable amount of lidocainehydrochloride.

Solutions for inhalation, injection or infusion may contain as carrier,for example, sterile water or preferably they may be in the form ofsterile, aqueous, isotonic saline solutions. Pharmaceutical compositionssuitable for delivery by needleless injection, for example,transdermally, may also be used.

A therapeutically or prophylactically effective amount of the compoundof the invention is administered to a subject. The dose may bedetermined according to various parameters, especially according to thecompound used; the age, weight and condition of the subject to betreated; the route of administration; and the required regimen. Again, aphysician will be able to determine the required route of administrationand dosage for any particular subject. A typical daily dose is fromabout 0.01 to 100 mg per kg, preferably from about 0.1 mg/kg to 50mg/kg, e.g. from about 1 to 10 mg/kg of body weight, according to theactivity of the specific inhibitor, the age, weight and conditions ofthe subject to be treated, the type and severity of the disease and thefrequency and route of administration. Preferably, daily dosage levelsare from 5 mg to 2 g.

When the compound of the invention is administered to a subject incombination with another active agent (for example in the form of apharmaceutical combination comprising an antibiotic agent and optionallyan SBL inhibitor), the dose of the other active agent (e.g. SBLinhibitor and/or antibiotic agent) can be determined as described above.The dose may be determined according to various parameters, especiallyaccording to the agent used; the age, weight and condition of thesubject to be treated; the route of administration; and the requiredregimen. Again, a physician will be able to determine the required routeof administration and dosage for any particular subject. A typical dailydose is from about 0.01 to 100 mg per kg, preferably from about 0.1mg/kg to 50 mg/kg, e.g. from about 1 to 10 mg/kg of body weight,according to the activity of the specific inhibitor, the age, weight andconditions of the subject to be treated, the type and severity of thedisease and the frequency and route of administration. Preferably, dailydosage levels are from 5 mg to 2 g.

The antibacterial properties of the compounds described herein mean thatthey are also useful in the treatment of bacterial infection in vitro,i.e. other than by the treatment of human or animal subjects. Thus, theinvention also provides a cleaning composition comprising a thiazolederivative of Formula (I) or a salt thereof. The cleaning compositionmay further comprise, for example, a detergent, a surfactant (includingionic and non-ionic surfactants), a diluent, a bleach (including ahypochlorite such as sodium hypochlorite or calcium hypochlorite,chlorine, chlorine dioxide, hydrogen peroxide or an adduct thereof,sodium perborate, and sodium percarbonate), an alcohol (such as ethanolor isopropanol), or a disinfectant. Typically, the disinfectant may beselected from benzyl-4-chlorophenol, amylphenol, phenylphenol,glutaraldehyde, alkyl dimethyl benzyl ammonium chloride, alkyl dimethylethylbenzyl ammonium chloride, iodine, peracetic acid and chlorinedioxide. Typically, the detergent may be an alkaline detergent such assodium hydroxide, sodium metasilicate, or sodium carbonate, or an aciddetergent such as hydrochloric acid, nitric acid, sulfuric acid,phosphoric acid, citric acid, or tartaric acid.

The following Examples illustrate the invention. They do not however,limit the invention in any way. In this regard, it is important tounderstand that the particular assay used in the Examples section isdesigned only to provide an indication of biological activity. There aremany assays available to determine biological activity, and a negativeresult in any one particular assay is therefore not determinative.

Experimental Details

General Synthetic Methodology

There are several related synthetic methods to this class of compoundsdescribed by Formula 1 and which are described below, where R is takento mean any substituent on the phenyl ring.

The preparation of the key thiazole intermediate tert-butyl5-{[(4-methoxyphenyl) methyl]amino}-1,3-thiazole-4-carboxylate has beendescribed previously (WO2014/198849) and is easily prepared on a 100 gscale. Reaction of this with a wide range of arylsulphonyl chlorides hasbeen achieved using basic catalysis (such as pyridine, triethylamine orsodium hydride) giving sulphonamide intermediates such as [A]. Otherversions of the thiazole starting material (eg R1=ethyl; R2=H) are alsoeasily accessible or even commercially available. Many of the compoundsdescribed herein are accessible from the standard Buchwald reaction ofbromophenylsulphonamide [A] with e.g. protected glycinamides such as[B]. Global acid-catalysed deprotection reveals the primary amine [C]which can be converted to the guanidine [D] if required (Scheme 1) usinga guanidinylating reagent such as 1H-pyrazole-1-carboximidamide.

Alternatively, instead of forming the aryl-nitrogen bond by usingBuchwald chemistry on the aryl bromide, it is possible to react certainaniline intermediates such as [E] with the N-protected glycine acidsusing standard peptide coupling reagents such as HATU, (Scheme 2).Deprotection and guanidinylation then gives [C] and [D] again,respectively. The anilines [E] are available from the correspondingnitro compounds by standard reduction or from the bromo intermediates bya Buchwald reaction using ammonia (eg see Scheme 4).

In certain circumstances, eg where the substituents on the aryl ring areparticularly electron-withdrawing, neither the Buchwald amidation northe amide formation using protected glycine derivatives are successful.For these situations it is necessary to react the aniline with highlyreactive chloroacetyl chloride to give intermediate [F]. Displacementwith sodium azide then affords azidoacetamide [G] which can be reducedwith standard reducing agents, accessing [C] and [D] in the usualmanner, (Scheme 3).

Certain ureido derivatives require bespoke syntheses (Scheme 4). Forexample Buchwald reaction of the usual bromoarylsulphonamide withammonia itself as the nitrogen-containing component gives thecorresponding aniline. Activation of this aniline with 4-nitrophenylchloroformate gives [H] which reacts with BOC-protected hydrazine [I] togive coupled product [J], possibly by the intermediacy of the isocyanatederived from [H]. Mild acid treatment removes the BOC group which can beguanidinylated to afford a protected guanidine functionality. Globaldeprotection of BOC, p-methoxybenzyl and tert-butyl ester groups thenaffords guanidine [K].

Certain analogues require a critical glyoxamide intermediate [M], whichis synthesised by reacting the usual aniline with 0.5 equivalents offumaryl chloride to give symmetrical bis amide [L]. Ozonolysis proceedsto give the labile glyoxamide [M] which can be reacted with a variety ofnucleophiles including the bis-BOC protected aminoguanidine affording[N]. Global deprotection in the usual way then affords the correspondingimine [O], (Scheme 5).

Abbreviations

ACN Acetonitrile

AcOH Acetic acid

Ag(OTf) Silver triflate

AIBN Azobisisobutyronitrile

Boc Tert-butoxy-carbonyle

Boc2O Di-tert-butyl dicarbonate

Cs₂CO₃ Cesium carbonate

CFU Colony forming unit

CuI Copper iodide

DCM Dichloromethane

DIPEA N,N-Diisopropylethylamine

DMAP 4-Dimethylaminopyridine

DMF Dimethylformamide

DMS Dimethylsulfide

DMSO Dimethyl sulfoxide

dppf 1,1′-Bis(diphenylphosphino)ferrocene

EDC.HCl N-(3-Dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride

EtOAc Ethyl acetate

EtOH Ethanol

Et3N Triethylamine

HATU 1-[Bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium3-oxid hexafluorophosphate

HCl Hydrochloric acid

HOBt Hydroxybenzotriazole

H2SO4 Sulfuric Acid

IPA Iso-propyl alcohol

Km Michaelis constant

MeI Methyl iodide

MeOH Methanol

NBS N-bromo succinimide

Na2CO3 Sodium carbonate

Na2SO4 Sodium sulfate

Pd₂(dba)₃ Tris(dibenzylideneacetone)dipalladium(O)

PdCl₂(PPh3)2 Bis(triphenylphosphine)palladium(II) dichloride

PdCl₂(dppf) [1,1′-Bis(diphenylphosphino)ferrocene]dichloropalladium(II)

PMB Paramethoxybenzyl

TEA Triethylamine

TES Triethylsilane

TMSOK Potassium trimethylsilanolate

TFA Trifluoroacetic acid

TMSOTf Trimethylsilyl trifluoromethanesulfonate

TFA Trifluoroacetic acid

THF Tetrahydrofurane

T3P Propylphosphinic anhydride

RT Room temperature

The structure of (RuPhos) Palladium (II) phenethylamine chloride (1:1MTBE adduct) used for Buchwald coupling steps (RuPhos Pd G1 complex) isshown below.

EXAMPLES

General Techniques

1H NMR spectra are reported at 300 or 400 MHz in DMSO-d6 solutions (δ inppm), using chloroform as the reference standard (7.25 ppm). When peakmultiplicities are reported, the following abbreviations are used: s(singlet), d (doublet), t (triplet), m (multiplet), bs (broadenedsinglet), dd (doublet of doublets), dt (doublet of triplets), q(quartet). Coupling constants, when given, are reported in hertz (Hz).

The term “purified by prep hplc (MDAP)” refers compound purificationusing a mass-directed auto purification system on an Agilent 1260infinity machine with an XSelect CHS Prep C18 column, eluting with 0.1%formic acid in water/acetonitrile and detection with a Quadruploe LC/MS.

Example 1 tert-butyl5-[(4-methoxyphenyl)methylamino]thiazole-4-carboxylate

(Key Intermediate-1)

A suspension of potassium tert-butoxide (874 mg, 7.79 mmol) in drytetrahydrofuran (10 mL) was stirred vigorously at room temperature. Tothis, a solution of tert-butyl isocyanoacetate (1.0 g, 7.08 mmol) in drytetrahydrofuran (5 mL) was added drop wise and the mixture stirred atroom temperature for 10 minutes. To this, a solution of 4-methoxybenzylisothiocyanate (1.27 g, 7.08 mmol) in dry tetrahydrofuran (5 mL) wasadded drop wise at room temperature. After 2 hours the solution waspoured into saturated NaHCO3 solution and extracted with ethyl acetate.The organic layer was dried with Na2SO4, filtered and concentrated invacuo to dryness. The residue was purified by silica gel chromatography(eluting with 0-50% ethyl acetate/cyclohexane) affording the titleproduct as a pale yellow solid (852 mg).

1H NMR (CDC₃) δ: 7.81 (1H, m), 7.73 (1H, br s), 7.31-7.23 (2H, m),6.92-6.85 (2H, m), 4.35 (2H, d), 3.80 (3H, s), 1.61 (9H, s).

M/z 321 (M+H)⁺

Example 25-[[3,5-difluoro-4-[(2-guanidinoacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylicacid

a. tert-butyl5-[(4-bromo-3,5-difluoro-phenyl)sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

A solution of tert-butyl5-[(4-methoxyphenyl)methylamino]thiazole-4-carboxylate (1 g, 3.12 mmol,1 eq) in THF (15 mL) was added to NaH suspension in THF (10 mL) at 0° C.under argon atmosphere. After 30 minutes, a solution of4-bromo-3,5-difluoro-benzenesulfonyl chloride (1.0 g, 3.43 mmol, 1.1 eq)in THF (15 mL) was added at 0° C. under argon atmosphere. The resultingreaction mixture was stirred at RT for 1 h, quenched with ice cold water(20 mL) and extracted with ethyl acetate (2×20 mL). The combined organiclayer was dried over Na₂SO₄, filtered and concentrated. The crudematerial was purified by trituration with diethyl ether (2×5 mL)affording a pale yellow solid (800 mg, 44%).

M/z 577.0 (M+H)⁺

b. tert-butyl5-[[4-[[2-(tert-butoxycarbonylamino)acetyl]amino]-3,5-difluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

A solution of tert-butyl5-[(4-bromo-3,5-difluoro-phenyl)sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(100 mg, 0.173 mmol, 1 eq) in 1,4-dioxane (5 mL) was purged with argonfor 15 minutes. Then tert-butyl N-(2-amino-2-oxo-ethyl)carbamate (45 mg,0.26 mmol, 1.5 eq), K₃PO₄ (110 mg, 0.521 mmol, 3 eq), Pd₂(dba)₃ (16 mg,0.17 mmol, 0.1 eq) and Xantphos (30 mg, 0.052 mmol, 0.3 eq) were addedunder argon atmosphere. The resulting reaction mixture was heated to 85°C. for 16 h in a closed vial. The temperature was allowed to cool to RT,and the reaction mixture was filtered through celite pad and the pad waswashed with EtOAc (2×5 mL). The organic layer was concentrated underreduced pressure. The resulting crude compound was dissolved in ethylacetate (25 mL), washed with water (10 mL) and brine solution (10 mL).The organic layer was dried over Na₂SO₄, filtered and concentrated. Thecrude material was purified by flash chromatography (eluting with 55%ethyl acetate in petroleum ether) affording a pale yellow solid (60 mg,51%).

M/z 669.5 (M+H)⁺

c.5-[[4-[(2-aminoacetyl)amino]-3,5-difluoro-phenyl]sulfonylamino]thiazole-4-carboxylicacid, trifluoroacetate

TFA (4 mL) was added to tert-butyl5-[[4-[[2-(tert-butoxycarbonylamino)acetyl]amino]-3,5-difluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(300 mg, 0.448 mmol, 1 eq) at RT and stirred for 4 h. TFA was evaporatedby reduced pressure. The resulting crude product was triturated withdiethyl ether (2×5 mL) and dried under high vacuum to afford a paleyellow solid (150 mg, 85%).

M/z 393.3 (M+H)⁺

d.5-[[3,5-difluoro-4-[(2-guanidinoacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylicacid

To a stirred solution of5-[[4-[(2-aminoacetyl)amino]-3,5-difluoro-phenyl]sulfonylamino]thiazole-4-carboxylicacid, trifluoroacetate (150 mg, 0.382 mmol, 1 eq) in DMF (5 mL) wasadded pyrazole-1-carboxamidine; hydrochloride (84 mg, 0.573 mmol, 1.5eq) and DIPEA (0.3 mL, 1.91 mmol, 5 eq) at RT. The resulting reactionmixture was stirred at RT for 16 h, and concentrated under reducedpressure. Water (5 mL) was added to the residue and the precipitate wasfiltered and washed with diethyl ether (2×5 mL). The crude product waspurified by preparative HPLC to afford the title compound as a whitesolid (47 mg, 28%).

¹H NMR (400 MHz, DMSO-d₆) δ 13.20 (1H, s), 10.14 (1H, brs), 8.12 (1H,s), 7.55 (1H, brs), 7.43 (2H, d, J=7.2 Hz), 7.35-7.10 (3H, brs), 4.12(2H, s).

M/z 434.9 (M+H)⁺

LC-MS Condition:

Column: Acquity BEH C18 (50 mm×2.1 mm, 1.7 um);

Mobile Phase: A: 0.05% Formic Acid in water;

B: 0.05% Formic Acid in ACN;

Time (min)/% B: 0/3, 0.4/3, 2/98, 3.4/98, 3.5/3, 4/3;

Column Temp: 35° C., Flow Rate: 0.6 mL/min

Prep. HPLC Condition:

Column: Symmetry C18 (300*19) mm, 7 u;

Mobile phase: (A) 0.1% Formic Acid (B) Acetonitrile;

Flow: 19 mL/min;

Gradient (T/% B): 0/5, 1/5, 8/20, 11/20, 11.02/99, 12/99, 12.1/5, 15/5;

Solubility: ACN+H₂O+THF+FA

Example 35-[[5-fluoro-6-[(2-guanidinoacetyl)amino]-3-pyridyl]sulfonylamino]thiazole-4-carboxylicacid

a. 5-fluoro-6-hydroxy-pyridine-3-sulfonylchloride

3-fluoropyridin-2-ol (2 g, 17.6 mmol) was added to chlorosulfonic acid(20 mL, 300.3 mmol) at 0° C. The reaction mixture was stirred at 160° C.for 2 h, cooled to RT and slowly poured into ice cold water (50 mL). Theaqueous layer was extracted with EtOAc (3×50 mL). The organic layer wasdried over Na₂SO₄, filtered and concentrated under reduced pressure. Thecrude compound was triturated with n-pentane (2×50 mL) to afford anoff-white solid (2.7 g, 72%).

M/z 212.11 (M+H)⁺

b. 6-chloro-5-fluoro-pyridine-3-sulfonylchloride

Thionyl chloride (5 mL, 68.9 mmol) was added to5-fluoro-6-hydroxy-pyridine-3-sulfonyl chloride (1 g, 4.73 mmol) intoluene (25 mL) at 0° C. DMF (0.2 mL) was then added slowly at 0° C. Thereaction mixture was refluxed for 3 h, cooled to RT and concentratedunder reduced pressure. The resulting crude material was co-distilledwith toluene (2×25 mL) to afford a pale yellow liquid which was used inthe next step without further purification (0.9 g, crude).

¹H NMR (400 MHz, DMSO-d₆) δ 8.83 (1H, m), 8.04 (1H, m).

c. tert-butyl5-[(6-chloro-5-fluoro-3-pyridyl)sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

A solution of tert-butyl5-[(4-methoxyphenyl)methylamino]thiazole-4-carboxylate (1.5 g, 4.68mmol) in THF (25 mL) was added to NaH (1.12 g, 46.8 mmol) suspension inTHF (10 mL) at 0° C. under argon atmosphere. After 15 minutes, asolution of 6-chloro-5-fluoro-pyridine-3-sulfonyl chloride (1.6 g, 7.0mmol) in THF (15 mL) was added to the above reaction mixture at 0° C.under argon atmosphere. The resulting reaction mixture was stirred at RTfor 0.5 h, quenched with ice cold water (20 mL) and extracted with ethylacetate (3×20 mL). The combined organic layer was dried over Na₂SO₄,filtered and concentrated. The crude compound was purified by flashchromatography (eluting with 10-15% ethyl acetate in petroleum ether) toafford a yellow oil (1.3 g, 54%).

M/z 514.27 (M+H)⁺

d. tert-butyl5-[[6-[[2-(tert-butoxycarbonylamino)acetyl]amino]-5-fluoro-3-pyridyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

A solution of tert-butyl5-[(6-chloro-5-fluoro-3-pyridyl)sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(150 mg, 0.29 mmol) in 1,4-dioxane (5 mL) was purged with argon for 20minutes. Then tert-butyl N-(2-amino-2-oxo-ethyl)carbamate (75 mg, 0.43mmol), Cs₂CO3 (282 mg, 0.87 mmol), Pd₂(dba)₃ (26 mg, 0.02 mmol) andXantphos (50 mg, 0.08 mmol) were added under argon atmosphere. Theresulting reaction mixture was heated to 70° C. for 0.5 h in a sealedtube, allowed to cool to RT, filtered through celite pad and the pad waswashed with ethyl acetate (2×3 mL). The organic layer was concentratedunder reduced pressure. The crude material was purified by flashchromatography (eluting with 50% ethyl acetate in petroleum ether) toafford a pale yellow solid (75 mg, 39%).

M/z 652.41 (M+H)⁺

e.5-[[6-[(2-aminoacetyl)amino]-5-fluoro-3-pyridyl]sulfonylamino]thiazole-4-carboxylicacid, trifluoacetate

TFA (1.5 mL) was added to a solution of tert-butyl5-[[6-[[2-(tert-butoxycarbonylamino)acetyl]amino]-5-fluoro-3-pyridyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(150 mg, 0.23 mmol) in DCM (2 mL) at 0° C., allowed to stir at RT for 18h and concentrated under reduced pressure. The residue was trituratedwith diethyl ether (2×2 mL), n-pentane (2×2 mL) and dried under highvacuum to afford an off white solid which was used in next step withoutfurther purification (60 mg, crude).

M/z 376.24 (M+H)⁺

f.5-[[5-fluoro-6-[(2-guanidinoacetyl)amino]-3-pyridyl]sulfonylamino]thiazole-4-carboxylicacid

Pyrazole-1-carboxamidine; hydrochloride (70 mg, 0.48 mmol) and DIPEA(0.27 mL, 1.6 mmol) were added to a stirred solution of5-[[6-[(2-aminoacetyl)amino]-5-fluoro-3-pyridyl]sulfonylamino]thiazole-4-carboxylicacid, trifluoroacetate (120 mg, 0.32 mmol) in DMF (2 mL) at RT. Theresulting reaction mixture was stirred at RT for 4 h, concentrated underreduced pressure and ice cold 1N HCl (2 mL) was added to the crudecompound and stirred for 10 minutes. The resulting precipitate wasfiltered, washed with diethyl ether (2×5 mL) and dried under highvacuum. The crude product was purified by preparative. HPLC affordingthe title product as an off white solid (25 mg, 18%).

¹H NMR (400 MHz, DMSO-d₆) δ 13.20 (1H, brs), 10.8 (1H, brs), 8.51 (1H,d, J=1.6 Hz), 8.13 (1H, s), 7.99 (1H, dd, J=9.6 Hz, J=1.6 Hz), 7.52 (1H,brs), 7.26 (3H, brs), 4.20 (2H, d, J=4.4 Hz).

M/z 418.18 (M+H)⁺

LC-MS Condition:

Column: Acquity BEH C18 (50 mm×2.1 mm, 1.7 um);

Mobile Phase: A: 0.05% Formic Acid in water; B: 0.05% Formic Acid inACN;

Time (min)/% B: 0/3, 0.4/3, 3.2/98, 3.8/98, 4.2/3, 4.5/3;

Column Temp: 35° C.;

Flow Rate: 0.6 mL/min.

Prep. HPLC Condition:

Column used: PHENYL HEXYL (150*30) mm 5 u;

Mobile phase: (A) 0.1% Formic Acid, (B) Acetonitrile;

Flow: 19 mL/min;

Gradient −(T/% B): 0/5, 1/5, 6/30, 8.9/30, 8.95/99, 11/99, 11.1/5, 14/5;

Solubility: ACN+THF.

Example 45-[[4-[(3-amino-3-imino-propanoyl)amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylicacid

a. tert-butylN-[3-amino-1-(tert-butoxycarbonylamino)-3-oxo-prop-1-enyl]carbamate

Saturated NaHCO₃ solution (10 mL was added to a stirred solution of3-amino-3-imino-propanamide (3 g, 29.6 mmol) in dioxane (20 mL) at RT.Then (Boc)₂O (16.5 mL, 74.0 mmol) was added drop wise at 0° C. Theresulting reaction mixture was stirred at RT for 16 h, concentratedunder reduced pressure and water (30 mL) was added to the residue. Thecrude compound was extracted with ethyl acetate (2×50 mL). The combinedorganic layer was dried over Na₂SO₄, filtered and concentrated. Thecrude material was purified by flash column chromatography (eluting with2% methanol in DCM) to afford an off-white solid (3.1 g, 34%).

M/z 302.36 (M+H)⁺

b. tert-butyl5-[[4-[3,3-bis(tert-butoxycarbonylamino)prop-2-enoylamino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

A solution of tert-butyl5-[(4-bromo-3-fluoro-phenyl)sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(1.1 g, 1.97 mmol) in 1,4-dioxane (15 mL) was purged with argon for 15minutes. Then tert-butylN-[3-amino-1-(tert-butoxycarbonylamino)-3-oxo-prop-1-enyl]carbamate (892mg, 2.95 mmol), K₃PO₄ (837 mg, 3.94 mmol), Pd₂(dba)₃ (180 mg, 0.19 mmol)and Xantphos (342 mg, 0.59 mmol) were added under argon atmosphere. Theresulting reaction mixture was heated to 65° C. for 3 h in a sealedtube, cooled to RT, filtered through a celite pad and the pad was washedwith EtOAc (2×10 mL). The filtrate was concentrated under reducedpressure. The resulting crude compound was dissolved in ethyl acetate(50 mL), washed with water (30 mL) and brine solution (30 mL). Theorganic layer was dried over Na₂SO₄, filtered and concentrated underreduced pressure. The crude material was purified by flashchromatography (eluting with 55% ethyl acetate in petroleum ether) toafford a pale yellow solid (1.3 g, 85%).

M/z 778.52 (M+H)⁺

c.5-[[4-[(3-amino-3-imino-propanoyl)amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylicacid

TFA (6 mL) was added to tert-butyl5-[[4-[3,3-bis(tert-butoxycarbonylamino)prop-2-enoylamino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(600 mg, 0.77 mmol) at RT. The resulting mixture was stirred for 3 h andconcentrated under reduced pressure. The resulting crude product wastriturated with diethyl ether (2×10 mL) and dried under high vacuum. Thecrude product was purified by preparative HPLC affording the titleproduct as an off-white solid (47 mg, 15%).

¹H NMR (400 MHz, DMSO-d₆) δ 13.42 (1H, brs), 10.34 (1H, brs), 8.99 (2H,brs), 8.62 (2H, brs), 8.14-8.02 (2H, m), 7.58-7.50 (2H, m), 3.68 (2H,s).

M/z 402.3 (M+H)⁺

LC-MS Condition:

Column: Acquity BEH C18 (50 mm×2.1 mm, 1.7 um);

Mobile Phase: A: 0.05% Formic Acid in water; B: 0.05% Formic Acid inACN;

Time (min)/% B: 0/3, 0.4/3, 3.2/98, 3.8/98, 4.2/3, 4.5/3;

Column Temp: 35° C.;

Flow Rate: 0.6 mL/min.

Prep. HPLC Condition:

Column used: PRONTOSIL (250*19) mm, 5 u;

Mobile phase: (A) 0.1% Formic Acid, (B) Acetonitrile;

Flow: 19 mL/min;

Gradient −(T/% B): 0/5, 1/5, 7.3/59, 7.4/99, 11/99, 11.1/5, 14/5;

Solubility: ACN+THF+H₂O+formic acid.

Examples 5 and 6 Example 5:5-[[3-cyano-4-[(2-guanidinoacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylicacid

Example 6:5-[[3-carbamoyl-4-[(2-guanidinoacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylicacid

a. 4-bromo-3-cyano-benzenesulfonylchloride

Solution-A: To a stirred solution of 5-amino-2-bromo-benzonitrile (2 g,10.1 mmol) in AcOH (25 mL) was added conc. HCl (5 mL) at 0° C. andstirred for 10 minutes. Then NaNO₂ (770 mg, 11.1 mmol) in H₂O (10 mL)was added at the same temperature and stirred for 20 minutes.

Solution-B: SO2 gas was purged in AcOH (25 mL) for 30 minutes. Then asolution of CuCl₂ (1.62 g, 12.2 mmol) in H₂O (10 mL) was added at 0° C.and stirred for 20 minutes. After that, Solution-B was added drop wiseto Solution-A. The reaction mixture was stirred at RT for 20 minutes anddiluted with water (20 mL). The resulting precipitate was filtered,washed with n-pentane (2×20 mL) and dried under high vacuum to afford ayellow solid (1.7 g, 60%).

b.5-[[3-cyano-4-[(2-hydroxyacetyl)amino]phenyl]sulfonyl-methyl-amino]thiazole-4-carboxylicacid

This compound was prepared following the procedure reported for Example2 step b.

M/z 658.8 (M+H)⁺

c.5-[[4-[(2-aminoacetyl)amino]-3,5-difluoro-phenyl]sulfonylamino]thiazole-4-carboxylicacid, trifluoroacetate

This compound was prepared following the procedure reported for Example2 step c.

M/z 382.4 (M+H)⁺

d.5-[[4-[(2-aminoacetyl)amino]-3-cyano-phenyl]sulfonylamino]thiazole-4-carboxylicacid and5-[[4-[(2-aminoacetyl)amino]-3-carbamoyl-phenyl]sulfonylamino]thiazole-4-carboxylicacid

TFA: H₂O (9:1, 5 mL) was added to tert-butyl5-[[4-[[2-(tert-butoxycarbonylamino)acetyl]amino]-3-cyano-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(400 mg, 0.60 mmol) at RT. The reaction mixture was stirred for 6 h andconcentrated under reduced pressure. The resulting material wastriturated with diethyl ether (2×10 mL) and dried under high vacuum toobtain a yellow solid which was used in the next step without furtherpurification (300 mg, crude) (72% of5-[[4-[(2-aminoacetyl)amino]-3-cyano-phenyl]sulfonylamino]thiazole-4-carboxylicacid and 8% of5-[[4-[(2-aminoacetyl)amino]-3-carbamoyl-phenyl]sulfonylamino]thiazole-4-carboxylicacid).

M/z 382.05 (M+H)⁺ and 400.01 (M+H)⁺

e.5-[[3-cyano-4-[(2-guanidinoacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylicacid and5-[[3-carbamoyl-4-[(2-guanidinoacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylicacid

Pyrazole-1-carboxamidine (172 mg, 1.18 mmol) and DIPEA (0.3 mL, 1.57mmol) were added to a stirred solution of5-[[4-[(2-aminoacetyl)amino]-3-cyano-phenyl]sulfonylamino]thiazole-4-carboxylicacid and5-[[4-[(2-aminoacetyl)amino]-3-carbamoyl-phenyl]sulfonylamino]thiazole-4-carboxylicacid (300 mg, 0.78 mmol) in DMF (5 mL) at RT. The resulting reactionmixture was stirred at RT for 16 h and concentrated under reducedpressure. Water (5 mL) was added to the residue. The resultingprecipitate was filtered and washed with diethyl ether (2×5 mL). Thecrude product was purified by preparative HPLC affording the titleproducts:

Example 5

(72 mg, off-white solid):

¹H NMR (400 MHz, DMSO-d₆) δ 13.30 (1H, brs), 10.50 (1H, brs), 8.09 (1H,s), 8.02 (1H, d, J=2.0 Hz), 7.99 (1H, dd, J=8.8 Hz, J=2.0 Hz), 7.82 (1H,d, J=8.8 Hz), 7.52 (2H, brs), 7.23 (3H, brs), 4.13 (2H, s).

M/z 424.34 (M+H)⁺

Example 6

(5.2 mg, off-white solid):

¹H NMR (400 MHz, DMSO-d₆) δ 13.42 (1H, brs), 12.0 (1H, brs), 8.59 (1H,brs), 8.51 (1H, d), 8.20 (1H, d, J=2.0 Hz), 8.03 (1H, s), 7.83 (1H, dd,J=8.8 Hz, J=2.0 Hz), 7.80 (1H, brs), 7.44 (4H, brs), 4.07 (2H, s).

M/z 442.34 (M+H)⁺

LC-MS Condition:

Column: Acquity BEH C18 (50 mm×2.1 mm, 1.7 um);

Mobile Phase: A: 0.05% Formic Acid in water; B: 0.05% Formic Acid inACN;

Time (min)/% B: 0/3, 0.4/3, 3.2/98, 3.8/98, 4.2/3, 4.5/3;

Column Temp: 35° C.;

Flow Rate: 0.6 mL/min.

Prep. HPLC Condition:

Column: Symmetry C18 (150*25) mm, 10 u;

Mobile phase: (A) 0.05% Formic Acid (B) Acetonitrile;

Flow: 19 mL/min;

Gradient (T/% B): 0/5, 1/5, 5/20, 10.5/24, 10.52/99, 12/99, 12.02/5,15/5;

Solubility: ACN+H₂O+THF+FA.

Compounds prepared using analogous methods to those described forExamples 2 to 6 and purified in a similar manner by preparative HPLC areshown in the Table below.

Example Structure Name, NMR and mass 7

5-[[3-fluoro-4-[(2- guanidinoacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylic acid M/z 415.2 (M − Na)⁻ ¹H NMR(d6-DMSO) δ 13.1 (1H, s), 8.08 (1H, s), 7.93 (1H, brs), 7.48 (2H, d, J =7 Hz), 3.88 (2H, s). 8

5-[[4-[[2-[carbamimidoyl(methyl) amino]acetyl]amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4- carboxylic acid M/z 431.2 (M + H)⁺ ¹HNMR (d6-DMSO) δ 13.30 (1H, s), 10.14 (1H, brs), 8.44 (1H, s), 8.13 (1H,m), 8.07 (1H, s), 7.57-7.32 (5H, brs), 4.27 (2H, s), 2.95 (3H, s). 9

5-[[2,5-difluoro-4-[(2-guanidinoacetyl)amino]phenyl]sulfonylamino]thiazole-4- carboxylic acid M/z 435.3 (M +H)⁺ ¹H NMR (d6-DMSO) δ 13.3 (1H, s), 10.3 (1H, brs), 8.1 (1H, s), 8.04(1H, m), 7.57 (2H, m), 7.39-7.09 (4H, brs), 4.10 (2H, s). 10

5-[[4-[[2-(4,5-dihydro-1H-imidazol-2-ylamino)acetyl]amino]-2,5-difluoro- phenyl]sulfonylamino]thiazole-4-carboxylic acid M/z 461.3 (M + H)+ ¹H NMR (d6-DMSO) δ 13.19 (1H, s),10.1 (1H, brs), 8.39 (2H, brs), 8.10 (1H, s), 8.03 (1H, m), 7.55 (1H,m), 4.09 (2H, s), 3.60 (4H, s). 11

5-[[4-[[(2R)-2-guanidinopropanoyl]amino]phenyl]sulfonylamino]thiazole-4- carboxylic acid M/z 413.3 (M +H)⁺ ¹H NMR (d6-DMSO) δ 13.6 (1H, s), 10.3 (1H, s), 8.03 (1H, s), 7.71(5H, m), 7.25- 6.96 (4H, brs), 4.25 (1H, t, J = 7.2 Hz), 1.40 (3H, d, J= 6.8 Hz). 12

5-[[4-[[3-(dimethylamino)-3-imino- propanyl]amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4- carboxylic acid M/z 430.3 (M + H)⁺ ¹HNMR (d6-DMSO) δ 13.5 (1H, brs), 9.51 (1H, brs), 8.45 (1H, t, J = 8 Hz),8.19 (1H, s), 7.91 (2H, m), 4.22 (2H, s), 3.42 (3H, s), 3.35 (3H, s). 13

5-[[4-[(3-amino-3-imino- propanoyl)amino]-3,5-difluoro-phenyl]sulfonylamino]thiazole-4- carboxylic acid M/z 420.3 (M + H)⁺ ¹HNMR (d6-DMSO) δ 13.21 (1H, s), 9.01 (2H, brs), 8.58 (2H, brs), 8.12 (1H,s), 7.43 (2H, d, J = 7.2 Hz), 3.62 (2H, s). 14

5-[[6-[(2-guanidinoacetyl)amino]pyridazin-3-yl]sulfonylamino]thiazole-4- carboxylic acid M/z 401.4 (M +H)⁺ ¹H NMR (d6-DMSO) δ 13.1 (1H, brs), 11.5 (1H, brs), 8.45 (1H, s),8.38 (1H, d, J = 9.6 Hz), 8.21 (1H, brs), 8.10 (1H, s), 8.07 (1H, m),7.89-7.68 (4H, brs), 4.13 (2H, s). Key intermediate 6-chloro-3-pyridazinesulfonyl chloride was prepared as described in the literature,K. Ashton et al, WO2013/123444. 15

5-[[4-[[2-(4,5-dihydro-1H-imidazol-2-ylamino)acetyl]amino]-3,5-difluoro- phenyl]sulfonylamino]thiazole-4-carboxylic acid M/z 461.3 (M + H)+ ¹H NMR (d6-DMSO) δ 13.1 (1H, brs),10.1 (1H, brs), 8.12 (1H, s), 7.42 (2H, d, J = 9.6 Hz), 4.14 (2H, s),3.59 (4H, s). 16

5-[[4-[(2-guanidinoacetyl)amino]-3-methoxy-phenyl]sulfonylamino]thiazole- 4-carboxylic acid M/z 429.3 (M +H)⁺ ¹H NMR (d6-DMSO) δ 13.59 (1H, brs), 9.41 (1H, brs), 8.44 (1H, s),8.10 (1H, d, J = 8.4 Hz), 8.04 (1H, s), 7.77 (1H, brs), 7.56-7.32 (4H,brs), 7.30 (2H, m), 4.09 (2H, s), 3.86 (3H, s). The key intermediate4-acetamido-3- methoxybenzenesulfonyl chloride was prepared byliterature procedures, P. Patel et al, Bioorg Med Chem Lett, 2007, 17,6610. 17

5-[[3-chloro-4-[(2-guanidinoacetyl)amino]phenyl]sulfonylamino]thiazole-4- carboxylic acid M/z 433.3 (M +H)⁺ ¹H NMR (d6-DMSO) δ 13.4 (1H, brs), 9.81 (1H, brs), 8.43 (1H, s),8.08 (1H, s), 7.95 (1H, d, J = 8.4 Hz), 7.38 (1H, d, J = 2 Hz), 7.67(1H, m), 7.51-7.22 (4H, brs), 4.12 (2H, s). The key intermediate3-chloro-4- nitrobenzene-1-sulfonyl chloride is commercially available.16

5-[[3-cyano-4-[(2- guanidinoacetyl)amino]phenyl]sulfonyl-amino]thiazole-4-carboxylic acid M/z 424.3 (M + H)⁺ ¹H NMR (d6-DMSO) δ13.26 (1H, brs), 10.51 (1H, brs), 8.09 (1H, s), 8.03 (1H, m), 7.99 (1H,m), 7.83 (1H, d, J = 8.8 Hz), 7.52 (1H, brs), 7.38-7.13 (4H, brs), 4.13(2H, s). 17

5-[[3-carbamoyl-4-[(2- guanidinoacetyl)amino]phenyl]sulfonyl-amino]thiazole-4-carboxylic acid M/z 442.3 (M + H)⁺ ¹H NMR (d6-DMSO) δ13.41 (1H, brs), 12.01 (1H, brs), 8.59 (1H, brs), 8.51 (1H, d, J = 8.8Hz), 8.41 (1H, s), 8.20 (1H, d, J = 2 Hz), 8.04 (1H, s), 7.84 (1H, m),7.82 (1H, m), 7.54-7.30 (4H, brs), 4.07 (2H, s). 18

5-[[4-[(2-guanidinoacetyl)amino]-3-(trifluoromethoxy)phenyl]sulfonylamino] thiazole-4-carboxylic acid M/z483.0 (M + H)⁺ ¹H NMR (d6-DMSO) δ 13.38 (1H, s), 10.1 (1H, brs), 8.17(1H, d, J = 9 Hz), 8.09 (1H, s), 7.72 (1H, m), 7.64 (1H, d, J = 1.5 Hz),7.54 (1H, brs), 7.39-7.25 (4H, brs), 4.12 (2H, s). Key intermediate4-bromo-3- (trifluoromethoxy)benzenesulfonyl chloride was preparedaccording to literature procedures, C-M. Park et al, J Med Chem, 2008,51, 6902 19

5-[[3-fluoro-4-(3- guanidinopropanoylamino)phenyl]sulfon-ylamino]thiazole-4-carboxylic acid M/z 431.2 (M + H)⁺ ¹H NMR (d6-DMSO) δ10.19 (1H, s), 8.37 (1H, s), 8.18 (1H, t, J = 8.1 Hz), 7.60 (2H, m),7.46 (1H, m), 7.35-6.81 (4H, brs), 3.40 (2H, m), 2.70 (2H, t, J = 6.3Hz).

Example 205-[[4-[(2-guanidinoacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylicacid

a. ethyl5-[[4-[[2-(tert-butoxycarbonylamino)acetyl]amino]phenyl]sulfonylamino]thiazole-4-carboxylate

DIPEA (0.63 mL, 3.66 mmol) and HATU (696 mg, 1.83 mmol) were added to astirred solution of 2-(tert-butoxycarbonylamino)acetic acid (321 mg,1.83 mmol) in DMF (5 mL). The reaction mixture was stirred at RT for 15minutes and then ethyl5-[(4-aminophenyl)sulfonylamino]thiazole-4-carboxylate (400 mg, 1.22mmol) was added at the same temperature under N₂ atmosphere. Theresulting reaction mixture was stirred at RT for 16 h and concentratedunder reduced pressure. The resulting crude compound was dissolved in10% MeOH in DCM (20 mL), washed with sat NH₄Cl (2×10 mL), water (10 mL)and brine solution (10 mL). The organic layer was dried over Na₂SO₄,filtered and concentrated under vacuum. The crude material was purifiedby column chromatography (eluting with 3% MeOH) affording an off-whitesolid (400 mg, 67%).

M/z 484.8 (M+H)⁺507.06 (M+Na)⁺

b. ethyl5-[[4-[(2-aminoacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylate

2N HCl in Et₂O (4 mL) was added to ethyl5-[[4-[[2-(tert-butoxycarbonylamino)acetyl]amino]phenyl]sulfonylamino]thiazole-4-carboxylate(400 mg, 0.82 mmol) in diethyl ether (5 mL) at 0° C. The reactionmixture was stirred for 5 h at RT and concentrated under reducedpressure. The crude product was purified by preparative HPLC(HCOOH/CH₃CN/H₂O) affording an off-white solid (300 mg, 94%).

M/z 385.13 (M+H)⁺

c. ethyl5-[[4-[[2-[[N,N′-bis(tert-butoxycarbonyl)carbamimidoyl]amino]acetyl]amino]phenyl]sulfonylamino]thiazole-4-carboxylate

DIPEA (0.08 mL, 0.49 mmol) and tert-butylN-[(tert-butoxycarbonylamino)-pyrazol-1-yl-methylene]carbamate (87 mg,0.28 mmol) were added to a stirred solution of ethyl5-[[4-[(2-aminoacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylate(270 mg, 0.70 mmol) in DMF (5 mL) at RT. The resulting reaction mixturewas stirred at RT for 16 h and concentrated under reduced pressure. Theresulting crude compound was dissolved in 10% MeOH in DCM (20 mL),washed with water (10 mL) and brine solution (10 mL). The organic layerwas dried over Na₂SO₄, filtered and concentrated under reduced pressure.The crude material was purified by column chromatography (eluting with4% MeOH in DCM) affording an off white solid (250 mg, 56%).

M/z 626.97 (M+H)⁺

d.5-[[4-[[2-[[(Z)—N,N′-bis(tert-butoxycarbonyl)carbamimidoyl]amino]acetyl]amino]phenyl]sulfonylamino]thiazole-4-carboxylic acid

TMSOK (69 mg, 0.54 mmol) was added to a stirred solution of ethyl5-[[4-[[2-[[N,N′-bis(tert-butoxycarbonyl)carbamimidoyl]amino]acetyl]amino]phenyl]sulfonylamino]thiazole-4-carboxylate(170 mg, 0.27 mmol) in THF (4 mL) at RT under N₂ atmosphere. Theresulting reaction mixture was stirred at 40° C. for 5 h andconcentrated under reduced pressure. The resulting crude product wastriturated with diethyl ether (2×5 mL) and dried under high vacuum. Theresidue was dissolved in water (5 mL) and acidified with 1N HCl(adjusted pH2). The resulting solid was filtered, washed with n-pentaneand dried under high vacuum to afford an off-white solid which was usedto next step without further purification (70 mg crude, 43%).

M/z 598.92 (M+H)⁺

e.5-[[4-[(2-guanidinoacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylicacid

2N HCl in Ether (1 mL) was added to5-[[4-[[2-[[(Z)—N,N′-bis(tert-butoxycarbonyl)carbamimidoyl]amino]acetyl]amino]phenyl]sulfonylamino]thiazole-4-carboxylicacid (70 mg, 0.11 mmol) in diethyl ether (2 mL) at 0° C. The reactionmixture was stirred at RT for 5 h and concentrated under reducedpressure. The crude product was purified by preparative HPLC to affordthe title product as an off white solid (11 mg, 23%).

¹H NMR (500 MHz, DMSO-d₆) δ 13.59 (1H, s), 10.42 (1H, brs), 8.02 (1H,s), 7.68-7.63 (5H, m), 7.42 (4H, brs), 4.02 (2H, s).

M/z 398.78 (M+H)⁺

Example 215-[[4-[[2-(4,5-dihydro-1H-imidazol-2-yl)acetyl]amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylicacid

a. tert-butyl5-[[4-[(2-cyanoacetyl)amino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

A solution of 2-cyanoacetic acid (86 mg, 1.01 mmol) and PCl₅ (210 mg,1.01 mmol) in DCM (20 mL) was heated to reflux for 30 minutes. Thereaction mixture temperature was cooled to RT and a solution oftert-butyl5-[(4-amino-3-fluoro-phenyl)sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(500 mg, 1.01 mmol) in DCM (30 mL) was added under nitrogen atmosphere.The resulting reaction mixture was heated to reflux for 2.5 h, cooled toRT, diluted with DCM (50 mL) and washed with aqueous NaHCO₃ solution (30mL), water (30 mL) and brine (30 mL) solution. The organic layer wasdried over Na₂SO₄, filtered and concentrated under reduced pressure. Thecrude material was purified by flash chromatography (eluting with 1-2%MeOH in DCM) to afford a pale yellow solid (180 mg, 31%).

M/z 561.43 (M+H)⁺

b. tert-butyl5-[[4-[[2-(4,5-dihydro-1H-imidazol-2-yl)acetyl]amino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

HCl gas was passed to a solution of tert-butyl5-[[4-[(2-cyanoacetyl)amino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(300 mg, 0.53 mmol) in ethanol:Et₂O (1:2, 30 mL) at 0° C. for 2 h. Theresulting reaction mixture was kept in refrigerator for 16 h. Then thevolatile components were evaporated under reduced pressure at 40° C. Theresidue was dissolved in ethanol (10 mL) and ethylene diamine (48 mg,0.80 mmol) was added at RT. The reaction mixture was stirred at RT for16 h and concentrated under reduced pressure. The resulting crudeproduct was triturated with diethyl ether (2×5 mL) and dried under highvacuum to afford a pale brown solid which was used in the next stepwithout further purification (330 mg, crude).

M/z 548.29 (M-Boc+H)⁺

c.5-[[4-[[2-(4,5-dihydro-1H-imidazol-2-yl)acetyl]amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylicacid

TFA (3 mL) was added to tert-butyl5-[[4-[[2-(4,5-dihydro-1H-imidazol-2-yl)acetyl]amino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(300 mg, 0.54 mmol) at RT. The reaction mixture was stirred at RT for 4h and concentrated under reduced pressure. The resulting crude productwas triturated with diethyl ether (2×5 mL) and dried under high vacuum.The crude product was purified by preparative HPLC to afford the titleproduct as an off-white solid (26 mg, 11%).

¹H NMR (400 MHz, DMSO-d₆) δ 13.50 (1H, brs), 10.30 (1H, brs), 8.29 (2H,brs), 8.08-8.04 (2H, m), 7.54-7.48 (2H, m), 3.40 (2H, s), 3.36-3.28 (2H,obs), 2.88-2.85 (2H, m).

M/z 428.37 (M+H)⁺

Example 225-[[3-fluoro-4-[[3-imino-3-(methylamino)propanoyl]amino]phenyl]sulfonylamino]thiazole-4-carboxylicacid

a. tert-butyl5-[[3-fluoro-4-[[3-[hydroxy(methyl)amino]-3-imino-propanoyl]amino]phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

MeNHOH.HCl (298 mg, 3.56 mmol) and sodium carbonate (472 mg, 4.45 mmol)were added to a solution of tert-butyl5-[[4-[(2-cyanoacetyl)amino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(1 g, 1.78 mmol) in ethanol (15 mL) at RT. The resulting reactionmixture was stirred at 60° C. for 3 h, cooled to RT, filtered and washedwith ethanol (2×10 mL). The combined organic layer was concentratedunder reduced pressure. The obtained crude compound was triturated withEt₂O (2×10 mL) and dried under high vacuum to afford a brown solid whichwas used in next step without further purification.

M/z 608.03 (M+H)⁺

b. tert-butyl5-[[3-fluoro-4-[[3-imino-3-(methylamino)propanoyl]amino]phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

Bis(pinacolato)diboron (Adv. Synth. catal. 2015, 357, 451-462) (357 mg,1.4 mmol) was added to a solution of tert-butyl5-[[3-fluoro-4-[[3-[hydroxy(methyl)amino]-3-imino-propanoyl]amino]phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(570 mg, 0.93 mmol) in acetonitrile (10 mL) at RT. The resultingreaction mixture was stirred at RT for 1 h and concentrated underreduced pressure. The crude compound was purified by flashchromatography (eluting with 2% triethyl amine in 10% methanol and DCM)to afford a pale yellow solid (130 mg, 23%).

M/z 592.05 (M+H)⁺

c.5-[[3-fluoro-4-[[3-imino-3-(methylamino)propanoyl]amino]phenyl]sulfonylamino]thiazole-4-carboxylicacid

TFA (3 mL) was added to tert-butyl5-[[3-fluoro-4-[[3-imino-3-(methylamino)propanoyl]amino]phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(130 mg, 0.21 mmol) at RT. The reaction mixture was stirred for 2 h atRT and concentrated under reduced pressure. The resulting crude productwas triturated with diethyl ether (2×5 mL) and dried under high vacuum.The crude product was purified by preparative HPLC affording the titleproduct as yellow solid (20 mg, 22%).

¹H NMR (400 MHz, CF₃COOD) δ 9.53 (1H, brs), 8.42 (1H, t, J=8.0 Hz), 8.20(1H, s), 7.92 (1H, d, J=8.8 Hz), 7.88 (1H, d, J=9.2 Hz), 4.08 (2H, s),3.18 (3H, s).

M/z 416.34 (M+H)⁺

LC-MS Condition:

Column: Acquity BEH C18 (50 mm×2.1 mm, 1.7 um);

Mobile Phase: A: 0.05% Formic Acid in water; B: 0.05% Formic Acid inACN;

Time (min)/% B: 0/3, 0.4/3, 3.2/98, 3.8/98, 4.2/3, 4.5/3;

Column Temp: 35° C.;

Flow Rate: 0.6 mL/min.

Prep. HPLC Condition:

Column: Symmetry C18 (300*19) mm, 7 u;

Mobile phase: (A) 0.1% Formic Acid (B) Acetonitrile;

Flow: 19 mL/min;

Gradient (T/% B): 0/5, 1/5, 8.9/40, 8.92/99, 12/99, 12.1/5, 15/5;

Solubility: ACN+H₂O+THF.

Example 235-[[2-[(2-guanidinoacetyl)amino]thiazol-5-yl]sulfonylamino]thiazole-4-carboxylicacid

a. 2-acetamidothiazole-5-sulfonylchloride

N-thiazol-2-ylacetamide (5 g, 35.2 mmol) was added portion wise to asolution of chlorosulfonic acid (11.7 mL, 176 mmol) at 0° C. Thereaction mixture was stirred at 100° C. for 4 h, cooled to RT and pouredinto ice cold water (100 mL). The resulting precipitate was filtered andwashed with water (20 mL). The precipitate was triturated with n-pentane(2×20 mL) and azeotroped with toluene to afford an off-white solid whichwas used in the next step without further purification (2 g crude, 23%).

M/z 241.23 (M+H)⁺

b. ethyl5-[(2-acetamidothiazol-5-yl)sulfonylamino]thiazole-4-carboxylate

A solution of ethyl 5-aminothiazole-4-carboxylate (300 mg, 1.74 mmol) inTHF (10 mL) was added to a stirred solution of NaH (250 mg, 10.4 mmol)in THF (10 mL) at 0° C. and stirred for 5 minutes. Then a solution of2-acetamidothiazole-5-sulfonyl chloride (502 mg, 2.0 mmol) in THF (10mL) was added to the reaction mixture at 0° C. The reaction mixture wasstirred at the same temperature for 1 h. Ice cold water (30 mL) wasadded to the reaction mixture which was then washed with EtOAc (2×15mL). The aqueous layer was acidified to pH 2.0 using 1N HCl andextracted with EtOAc (3×15 mL). The organic layer was dried over Na₂SO₄,filtered and concentrated under reduced pressure to afford a pale brownsolid which was used to next step without further purification (175 mgcrude, 26%).

M/z 377.32 (M+H)⁺

c. ethyl 5-[(2-aminothiazol-5-yl)sulfonylamino]thiazole-4-carboxylate,hydrochloride

Concentrated HCl (7 mL) was added to a solution of ethyl5-[(2-acetamidothiazol-5-yl)sulfonylamino]thiazole-4-carboxylate (700mg, 1.86 mmol) in ethanol (70 mL) at RT. The reaction mixture wasrefluxed for 5 h and concentrated under reduced pressure. The resultingcrude compound was washed with diethyl ether (20 mL), n-pentane (20 mL)and dried under high vacuum to afford a brown solid which was used inthe next step without further purification (600 mg, crude).

M/z 335.04 (M+H)⁺

d. ethyl5-[[2-[[2-(tert-butoxycarbonylamino)acetyl]amino]thiazol-5-yl]sulfonylamino]thiazole-4-carboxylate

HATU (1.36 g, 3.58 mmol) and DIPEA (2.5 mL, 14.3 mmol) were added to astirred solution of 2-(tert-butoxycarbonylamino)acetic acid (628 mg,3.58 mmol) in DMF (6 mL) at RT. The reaction mixture was stirred at RTfor 15 minutes and then ethyl5-[(2-aminothiazol-5-yl)sulfonylamino]thiazole-4-carboxylate,hydrochloride (600 mg, 1.79 mmol) was added at the same temperatureunder N₂ atmosphere. The resulting reaction mixture was stirred at RTfor 18 h. Ice cold water (30 mL) was added and extracted with DCM (3×20mL). The organic layer was dried over Na₂SO₄, filtered and concentratedunder reduced pressure. The crude compound was purified by flashchromatography (eluting with using 60-80% of EtOAc in petroleum ether)affording a brown solid (400 mg, 45%).

M/z 492.34 (M+H)⁺

e.5-[[2-[[2-(tert-butoxycarbonylamino)acetyl]amino]thiazol-5-yl]sulfonylamino]thiazole-4-carboxylicacid

TMSOK (625 mg, 4.8 mmol) was added to a stirred solution of ethyl5-[[2-[[2-(tert-butoxycarbonylamino)acetyl]amino]thiazol-5-yl]sulfonylamino]thiazole-4-carboxylate(400 mg, 0.8 mmol) in THF (40 mL) at RT. The reaction mixture wasstirred at 40° C. for 1 h, concentrated under reduced pressure and water(2 mL) was added to the residue. The reaction mixture was acidified topH 2 using 1N HCl. The resulting precipitate was filtered, washed withdiethyl ether (2×10 mL), n-pentane (10 mL) and dried under high vacuumto afford a pale yellow solid (200 mg, 53%).

M/z 464.30 (M+H)⁺

f.5-[[2-[(2-aminoacetyl)amino]thiazol-5-yl]sulfonylamino]thiazole-4-carboxylicacid, hydrochloride

HCl in Et₂O (2M, 10 mL) was added to5-[[2-[[2-(tert-butoxycarbonylamino)acetyl]amino]thiazol-5-yl]sulfonylamino]thiazole-4-carboxylicacid (200 mg, 0.43 mmol) at RT. The reaction mixture was stirred at thesame temperature for 3 h, concentrated under reduced pressure and theresulting residue was washed with diethyl ether (2×5 mL) and n-pentane(5 mL) affording a pale yellow solid which was used in the next stepwithout further purification (150 mg, crude).

M/z 364.30 (M+H)⁺

g.5-[[2-[(2-guanidinoacetyl)amino]thiazol-5-yl]sulfonylamino]thiazole-4-carboxylicacid

DIPEA (0.44 mL, 2.7 mmol) was added to a stirred solution of5-[[2-[(2-aminoacetyl)amino]thiazol-5-yl]sulfonylamino]thiazole-4-carboxylicacid, hydrochloride (100 mg, 0.27 mmol) and pyrazole-1-carboxamidine,hydrochloride (80 mg, 0.55 mmol) in DMF (2 mL) at RT. The reactionmixture was stirred at the same temperature for 6 h. DMF was evaporatedand then water (3 mL) was added to the resulting crude material,stirring for 5 minutes. The resulting precipitate was filtered andwashed with water (2×2 mL) then dried under high vacuum. The crudecompound was purified by preparative HPLC affording an off-white solid(16 mg, 14%).

¹H NMR (400 MHz, DMSO-d₆) δ 13.28 (1H, s), 12.6 (1H, brs), 8.15 (1H, s),7.71 (1H, s), 7.44 (1H, t, J=6.4 Hz), 7.21 (4H, brs), 4.11 (2H, d, J=6.4Hz).

M/z 405.9 (M+H)⁺

LC-MS Condition:

Column: Acquity BEH C18 (50 mm×2.1 mm, 1.7 um)

Mobile Phase: A: 0.05% Formic Acid in water; B: 0.05% Formic Acid in ACN

Gradient: Time (min)/% B: 0/3, 0.4/3, 3.2/98, 3.8/98, 4.2/3, 4.5/3

Column Temp: 35° C.,

Flow Rate: 0.6 mL/min

Prep. HPLC Condition:

Column used: Atlantis T3 (250*19) mm, 5 u;

Mobile phase: (A) 0.1% Formic Acid (B) Acetonitrile

Flow: 19 mL/min

Gradient −(T/% B): 0/5, 1/5, 8.2/55, 8.21/99, 10/99, 10.1/5, 13/5

Diluent: ACN+H₂O+FA

Example 245-[[4-[2-(2-carbamimidoylhydrazino)-2-oxo-ethyl]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylicacid

a. tert-butyl5-[[4-(2-ethoxy-2-oxo-ethyl)-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

A mixture of tert-butyl5-[(4-bromo-3-fluoro-phenyl)sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(2 g, 3.58 mmol), potassium 3-ethoxy-3-oxo-propanoate (1.2 g, 7.16 mmol)and DMAP (43 mg, 0.35 mmol) in mesitylene (20 mL) was purged with argongas for 30 minutes. BINAP (222 mg, 0.35 mmol) and Pd2(dba)3 (327 mg,0.35 mmol) were then added at the same temperature. The reaction mixturewas stirred at 120° C. for 18 h, cooled to RT and concentrated underreduced pressure. The crude product was purified by flash chromatography(eluting with 40% EtOAc in petroleum ether) to afford a yellow solidwhich was used in the next step without further purification (0.45 g,crude).

M/z 565.43 (M+H)⁺

b. tert-butyl5-[[3-fluoro-4-(2-hydrazino-2-oxo-ethyl)phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

Hydrazine hydrate (709 mg, 14.1 mmol) was added to a stirred solution oftert-butyl5-[[4-(2-ethoxy-2-oxo-ethyl)-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(400 mg, 0.7 mmol) in ethanol (20 mL) at RT. The reaction mixture wasrefluxed for 5 h and concentrated under reduced pressure to afford abrown solid which was used in the next step without further purification(350 mg, crude).

M/z 551.42 (M+H)⁺

c. tert-butyl5-[[4-[2-[2-N,N′-bis(tert-butoxycarbonyl)carbamimidoyl]hydrazino]-2-oxo-ethyl]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

DIPEA (0.32 mL, 1.89 mmol) was added to a stirred solution of tert-butyl5-[[3-fluoro-4-(2-hydrazino-2-oxo-ethyl)phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(350 mg, 0.63 mmol) and tert-butylN—N-[(tert-butoxycarbonylamino)-pyrazol-1-yl-methylene]carbamate (394mg, 1.27 mmol) in DMF (5 mL) at RT. The reaction mixture was stirred atthe same temperature for 18 h. Ice cold water was added to the reactionmixture and stirred for 10 minutes. The resulting precipitate wasfiltered, washed with water (2×5 mL) and dried under high vacuum. Thecrude product was purified by flash chromatography (eluting with 60%EtOAc in petroleum ether) to afford a yellow solid (120 mg, 23%).

M/z 793.53 (M+H)⁺

d.5-[[4-[2-(2-carbamimidoylhydrazino)-2-oxo-ethyl]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylicacid

TFA (2 mL) was added to tert-butyl5-[[4-[2-[2-[(Z)—N,N′-bis(tert-butoxycarbonyl)carbamimidoyl]hydrazino]-2-oxo-ethyl]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(120 mg, 0.15 mmol) at RT. The reaction mixture was stirred for 3 h atthe same temperature and concentrated under reduced pressure. Theresulting crude material was triturated with diethyl ether (2×5 mL). Thecrude product was purified by preparative HPLC affording the titleproduct as an off-white solid (23 mg).

¹H NMR (400 MHz, DMSO-d6) δ 13.40 (1H, brs), 8.06 (1H, s), 7.63 (3H,brs), 7.51-7.39 (3H, m), 3.56 (2H, s).

M/z 417.35 (M+H)⁺

LC-MS Condition:

Column: Acquity BEH C18 (50 mm×2.1 mm, 1.7 um);

Mobile Phase: A: 0.05% Formic Acid in water; B: 0.05% Formic Acid inACN;

Time (min)/% B: 0/3, 0.4/3, 3.2/98, 3.8/98, 4.2/3, 4.5/3;

Column Temp: 35° C.;

Flow Rate: 0.6 mL/min.

Prep. HPLC Condition:

Column used: Symmetry C18 (300*19) mm, 7 u;

Mobile phase: (A) 0.05% Formic Acid (B) Acetonitrile;

Flow: 19 mL/min;

Gradient (T/% B): 0/2, 1/2, 8/20, 10.5/20, 10.51/99, 12/99, 12.1/2,15/2;

Solubility: ACN+H₂O+THF.

Compounds prepared by analogous methods to those described above forExamples 20 to 24 and purified in a similar manner by preparative HPLCare shown in the Table below:—

Example Structure Name, NMR and Mass 25

5-[[4-[2-[(2-amino-2-imino-ethyl)amino]-2-oxo-ethyl]-3-fluoro-phenyl]sulfonylamino] thiazole-4-carboxylic acid M/z416.1 (M + H)⁺ ¹H NMR (d6-DMSO) δ 13.1 (1H, s), 8.67 (1H, brs), 8.49(3H, brs), 8.07 (1H, brs), 7.48 (1H, m), 7.43 (2H, m), 3.89 (2H, s),3.61 (2H, s), 2.07 (1H, s).

Example 265-[[3,5-difluoro-4-(guanidinocarbamoylamino)phenyl]sulfonylamino]thiazole-4-carboxylicacid

a. tert-butyl5-[(4-amino-3,5-difluoro-phenyl)sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

A saturated solution of NH₃ in dioxane (120 mL) was added to a mixtureof tert-butyl5-[(4-bromo-3,5-difluoro-phenyl)sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(2 g, 3.47 mmol), Xantphos (0.6 g, 1.04 mmol), Pd₂(dba)₃ (0.317 g, 0.34mmol) and K₃PO₄ (2.2 g, 10.4 mmol). The resulting mixture was stirred insealed tube at 100° C. for 5 h, filtered through Celite pad and the padwas washed with ethyl acetate (2×25 mL). The filtrate was concentratedunder reduced pressure. The crude material was purified by flashchromatography (eluting with 50% ethyl acetate in petroleum ether)affording a pale yellow solid (1.25 g, 70%).

M/z 512.4 (M+H)⁺; 534.56 (M+Na)⁺

b. tert-butyl5-[[3,5-difluoro-4-[(4-nitrophenoxy)carbonylamino]phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

(4-nitrophenyl) carbonochloridate (1.57 g, 7.82 mmol) was added to astirred solution tert-butyl5-[(4-amino-3,5-difluoro-phenyl)sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(2 g, 3.91 mmol) in toluene (120 mL) at room temperature and refluxedfor 3 h. The reaction mixture was concentrated under reduced pressure.The crude product was used in the next step without further purification(3.5 g, crude).

c. tert-butyl5-[[4-[(tert-butoxycarbonylamino)carbamoylamino]-3,5-difluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

DIPEA (2.6 mL, 15.5 mmol) was added to a suspension of tert-butyl5-[[3,5-difluoro-4-[(4-nitrophenoxy)carbonylamino]phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(3.5 g, 5.17 mmol) and tert-butyl N-aminocarbamate (1.36 g, 10.3 mmol)in THF (100 mL) at 0° C. The reaction mixture was stirred at RT for 3 hand concentrated under reduced pressure. The crude material was purifiedby flash chromatography (eluting with 70% ethyl acetate in petroleumether) affording a pale yellow solid (1.5 g, 43%).

M/z 670.4 (M+H)⁺

d.5-[[3,5-difluoro-4-(hydrazinecarbonylamino)phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylicacid, hydrochloride

HCl in Et₂O (2M, 200 mL) was added to tert-butyl5-[[4-[(tert-butoxycarbonylamino)carbamoylamino]-3,5-difluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(1.5 g, 2.24 mmol) at RT. The reaction mixture was stirred for 24 h,cooled to 0° C. for 30 minutes and Et₂O was decanted. The crude productwas triturated with diethyl ether (2×40 mL) and dried under high vacuumaffording an off-white solid (1 g, crude).

M/z 514.3 (M+H)⁺

e.5-[[4-[[[N,N′-bis(tert-butoxycarbonyl)carbamimidoyl]amino]carbamoylamino]-3,5-difluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylicacid

DIPEA (3.0 mL, 17.5 mmol) was added to a stirred solution of5-[[3,5-difluoro-4-(hydrazinecarbonylamino)phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylicacid, hydrochloride (1 g, 1.75 mmol) and tert-butyl(NZ)—N-[(tert-butoxycarbonylamino)-pyrazol-1-yl-methylene]carbamate(0.54 g, 1.75 mmol) in DMF (6 mL) at RT. The reaction mixture wasstirred for 5 h and DMF was removed. Then water was added to the crudeproduct, stirring for 5 minutes. The resulting precipitate was filtered,washed with water (2×5 mL) and dried under high vacuum affording anoff-white solid (1.25 g, crude).

M/z 756.1 (M+H)⁺

f.5-[[3,5-difluoro-4-(guanidinocarbamoylamino)phenyl]sulfonylamino]thiazole-4-carboxylicacid

TFA (13 mL) was added to5-[[4-[[[(Z)—N,N′-bis(tert-butoxycarbonyl)carbamimidoyl]amino]carbamoylamino]-3,5-difluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylicacid (1.25 g, 1.54 mmol) at RT. The reaction mixture was stirred for 3 hat RT and TFA was evaporated by flushing with N₂ gas. The resultingcrude product was triturated with diethyl ether and purified bypreparative HPLC to afford the title compound as a white solid (150 mg).

¹H NMR (300 MHz, DMSO-d₆) δ 13.28 (1H, brs), 8.70 (3H, br s), 8.12 (1H,s), 7.39 (2H, d, J=6.9 Hz), 7.0-7.37 (3H, br s).

M/z 436.0 (M+H)⁺

LC-MS Condition:

Column: Aquity UPLC BEH C18 (50 mm×2.1 mm, 1.7 um);

Mobile phase: A: 0.1% of Formic Acid in Water, B: 0.1% of Formic acid inAcetonitrile;

Gradient: Time (min)/% B 0/2, 0.2/2, 1.5/98, 2.6/98, 2.61/2, 3.2/2;

Column Temp: 45° C., Flow rate: 0.8 mL/min

Prep. HPLC Condition:

Column: X select C18 (150*30 mm), 5 u;

Mobile Phase: 0.05% Formic acid in H₂O: Acetonitrile;

Flow: 25 mL/min;

Gradient (T/% B): 0/50, 8/50, 8/40, 9/40, 9.1/98, 11/98, 11.1/5, 14/40

Diluent: ACN+H₂O+MeOH+THF.

Example 275-[[3-fluoro-4-(2-guanidinoethoxycarbonylamino)phenyl]sulfonylamino]thiazole-4-carboxylicacid

a. tert-butyl5-[[3-fluoro-4-[(4-nitrophenoxy)carbonylamino]phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

p-Nitrophenyl chloroformate (1.33 g, 6.0 mmol) was added to a stirredsolution of tert-butyl5-[(4-amino-3-fluoro-phenyl)sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(1 g, 2.02 mmol) in toluene (30 mL) at RT. The resulting reactionmixture was stirred at 120° C. for 1 h and concentrated under reducedpressure. The resulting crude product was triturated with n-pentane(2×10 mL) and dried under high vacuum affording an off-white solid whichwas used in the next step without further purification (1.5 g, crude).

M/z 659.43 (M+H)⁺

b. tert-butyl5-[[4-[2-(tert-butoxycarbonylamino)ethoxycarbonylamino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

Tert-butyl N-(2-hydroxyethyl)carbamate (440 mg, 2.73 mmol) and DIPEA(0.97 mL, 5.46 mmol) were added to a stirred solution of tert-butyl5-[[3-fluoro-4-[(4-nitrophenoxy)carbonylamino]phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(1.2 g, 1.82 mmol) in THF (20 mL) at RT. The resulting reaction mixturewas stirred at RT for 2 h. Ice cold water was added followed byextraction with ethyl acetate (2×50 mL). The combined organic layer wasdried over anhydrous Na₂SO₄, filtered and concentrated under reducedpressure. The crude material was purified by flash chromatography(eluting with 40% ethyl acetate in petroleum ether) affording anoff-white solid (500 mg, 40%).

M/z 681.50 (M+H)⁺

c.5-[[4-(2-aminoethoxycarbonylamino)-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylicacid, trifluoroacetate

TFA (5 mL) was added to tert-butyl5-[[4-[2-(tert-butoxycarbonylamino)ethoxycarbonylamino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(450 mg, 0.66 mmol) at RT. The reaction mixture was stirred for 24 h atthe same temperature and concentrated under reduced pressure. Theresulting crude product was triturated with diethyl ether (2×10 mL) anddried under high vacuum to afford an off-white solid which was used inthe next step without further purification (350 mg, crude).

M/z 405.36 (M+H)⁺

d.5-[[3-fluoro-4-(2-guanidinoethoxycarbonylamino)phenyl]sulfonylamino]thiazole-4-carboxylicacid

Pyrazole-1-carboxamidine, hydrochloride (136 mg, 0.92 mmol) and DIPEA(0.55 mL, 3.0 mmol) were added to a stirred solution of5-[[4-(2-aminoethoxycarbonylamino)-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylicacid, trifluoroacetate (250 mg, 0.61 mmol) in DMF (6 mL) at RT. Theresulting reaction mixture was stirred at RT for 4 h concentrated underreduced pressure and water (5 mL) was added to the residue. Theresulting precipitate was filtered, washed with diethyl ether (2×5 mL)and dried under high vacuum. The crude product was purified bypreparative HPLC affording the title product as an off-white solid (45mg, 16%).

¹H NMR (300 MHz, DMSO-d₆) δ 13.42 (1H, brs), 9.60 (1H, s), 8.07 (1H, s),7.79 (1H, t, J=8.0 Hz), 7.62-7.56 (1H, m), 7.51 (1H, d, J=8.0 Hz, J=2.0Hz), 7.46 (1H, dd, J=10.4 Hz, 2.0 Hz), 7.12 (4H, brs), 4.18 (2H, t,J=5.2 Hz), 3.48-3.40 (2H, m).

M/z 447.27 (M+H)⁺

Compounds prepared using analogous methods to those described forExamples 26 and 27 and purified in a similar manner by preparative HPLCare shown in the Table below:

Example Structure Name, NMR and Mass 28

5-[[4-(carbamimidoylcarbamoyl-amino)-3- fluoro-phenyl]sulfonylamino]thiazole-4-carboxylic acid M/z 402.9 (M + H)⁺ ¹H NMR (d6-DMSO) δ 13.5(1H, brs), 8.18 (1H, brs), 8.13 (1H, m), 8.04 (1H, s), 7.72 (1H, brs),7.40 (2H, m), 6.96-6.61 (4H, m). 29

5-[[3-fluoro-4-(guanidinocarbamoylamino)phenyl]sulfonylamino]thiazole-4-carboxylic acid M/z 416.3 (M + H)⁺ ¹HNMR (d6-DMSO) δ 8.49 (1H, s), 8.04 (1H, brs), 7.88 (1H, brs), 7.50 (4H,m), 3.94 (2H, brs). 30

5-[[4-[(2-amino-2-imino-ethyl) carbamoylamino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylic acid M/z 417.3 (M + H)⁺ ¹H NMR(d6-DMSO) δ 13.5 (1H, brs), 9.01 (1H, s), 9.21-8.71 (4H, m), 8.43 (1H,s), 8.19 (1H, t, J = 8.4 Hz), 8.06 (1H, s), 7.46 (2H, t, J = 8.8 Hz),7.29 (1H, m), 4.04 (2H, d, J = 5.2 Hz). 31

5-[[3-fluoro-4-(2-guanidinoethylsulfanyl-carbonylamino)phenyl]sulfonylamino]thiazole- 4-carboxylic acid M/z 463.1(M + H)⁺ ¹H NMR (d6-DMSO) δ 13.42 (1H, s), 10.51 (1H, s), 8.16 (1H, s),8.08 (1H, s), 7.79 (1H, t, J = 7.5 Hz), 7.61 (1H, s), 7.51 (2H, m),7.31-6.79 (4H, m), 3.36 (2H, t, J = 6.5 Hz), 3.36 (2H, t, J = 6.5 Hz).

Example 325-[[3,5-difluoro-4-(guanidinocarbamoylamino)phenyl]sulfonylamino]thiazole-4-carboxylicacid

a. tert-butyl5-[[4-[(2-chloroacetyl)amino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

Et₃N (276 mg, 2.73 mmol) and chloroacetyl chloride (185 mg, 1.64 mmol)were added to a stirred solution of tert-butyl5-[(4-amino-3-fluoro-phenyl)sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(450 mg, 0.91 mmol) in DCM (5 mL) at 0° C. The resulting reactionmixture was stirred at RT for 2 h, quenched with ice cold water (5 mL)and extracted with DCM (2×10 mL). The combined organic layer was driedover Na2SO₄, filtered and concentrated under reduced pressure. The crudematerial was purified by trituration with diethyl ether (2×5 mL) toafford a green solid which was used in the next step without furtherpurification (400 mg, crude).

M/z 570.69 (M+H)⁺

b. tert-butyl5-[[4-[(2-azidoacetyl)amino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

NaN₃ (92 mg, 1.40 mmol) was added to a stirred solution of tert-butyl5-[[4-[(2-chloroacetyl)amino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]amino]thiazole-4-carboxylate(400 mg, 0.70 mmol) in DMF (5 mL) at RT. The resulting reaction mixturewas stirred at RT for 16 h, quenched with ice cold water (10 mL) andextracted with ethyl acetate (2×10 mL). The combined organic extractswere dried over Na₂SO₄, filtered and concentrated under reducedpressure. The crude material was purified by trituration with diethylether (2×5 mL) to afford a light brown solid (370 mg, 91%).

M/z 577.23 (M+H)⁺

c. tert-butyl5-[[4-[(2-aminoacetyl)amino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

10% Pd/C (300 mg) was added to solution of tert-butyl5-[[4-[(2-azidoacetyl)amino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(370 mg, 0.64 mmol) in EtOAc (10 mL) at RT under nitrogen atmosphere.The resulting reaction mixture was stirred at RT under a hydrogenatmosphere (balloon pressure) for 16 h, filtered through a pad of celiteand washing with EtOAc (20 mL). The filtrate was concentrated underreduced pressure. The crude material was purified by trituration withdiethyl ether (2×5 mL) to afford a brown solid (340 mg, 96%).

M/z 551.35 (M+H)⁺

d. tert-butyl5-[[4-[[2-(4,5-dihydro-1H-imidazol-2-ylamino)acetyl]amino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

2-Methylsulfanyl-4,5-dihydro-1H-imidazole (124 mg, 0.50 mmol) was addedto a stirred solution of tert-butyl5-[[4-[(2-aminoacetyl)amino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(400 mg, 0.72 mmol) in THF (5 mL) at RT. The resulting reaction mixturewas heated to 70° C. for 48 h in a closed vial and concentrated underreduced pressure to afford a yellow solid which was used in the nextstep without further purification (500 mg, crude).

M/z 619.36 (M+H)⁺

e.5-[[3,5-difluoro-4-(guanidinocarbamoylamino)phenyl]sulfonylamino]thiazole-4-carboxylicacid

TFA (5 mL) was added to tert-butyl5-[[4-[[2-(4,5-dihydro-1H-imidazol-2-ylamino)acetyl]amino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(500 mg, 0.50 mmol) at 0° C. and stirred at RT for 6 h. TFA wasevaporated by reduced pressure and the resulting crude product wastriturated with diethyl ether (2×5 mL) and dried under vacuum. The crudeproduct was purified by preparative HPLC affording the title product asa white solid (37 mg, 18%).

¹H NMR (400 MHz, DMSO-d₆) δ 13.43 (1H, brs), 10.20 (1H, brs), 8.4 (3H,brs), 8.11-8.08 (2H, m), 7.55-7.48 (2H, m), 4.08 (2H, s), 3.60 (4H, s).

M/z 443.24 (M+H)⁺

LC-MS Condition:

Column: Acquity BEH C18 (50 mm×2.1 mm, 1.7 um);

Mobile Phase: A: 0.05% Formic Acid in water; B: 0.05% Formic Acid inACN;

Time (min)/% B: 0/3, 0.4/3, 2/98, 3.8/98, 4.2/3, 4.5/3;

Column Temp: 35° C.,

Flow Rate: 0.6 mL/min.

Prep. HPLC Condition:

Column: Atlantis T3 (250*19) mm, 5 u;

Mobile phase: (A) 0.1% Formic Acid (B) Acetonitrile;

Flow: 19 mL/min;

Gradient (T/% B): 0/5, 1/5, 9/30, 10.31/99, 12/99, 12.1/5, 15/5;

Solubility: ACN+H₂O+THF.

Example 335-[[3-fluoro-4-[[2-(morpholine-4-carboximidoylamino)acetyl]amino]phenyl]sulfonylamino]thiazole-4-carboxylicacid

a. tert-butyl5-[[3-fluoro-4-[[2-(morpholine-4-carbothioylamino)acetyl]amino]phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

A solution of tert-butyl5-[[4-[(2-aminoacetyl)amino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(500 mg, 0.9 mmol) and di(imidazol-1-yl)methanethione (242 mg, 1.36mmol) in CH₂Cl₂ (15 mL) was stirred at RT for 30 minutes. Thenmorpholine (118 mg, 1.36 mmol) was added and the resulting reactionmixture was stirred at 40° C. for 1 h. The reaction mixture wasconcentrated under vacuum.

The crude compound was purified by flash chromatography eluting with 3%MeOH in CH₂Cl₂ to afford a pale pink gummy material (100 mg, 83%).

M/z 680.42 (M+H)⁺

b. tert-butyl5-[[3-fluoro-4-[[2-(morpholine-4-carboximidoylamino)acetyl]amino]phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

Ag(OTf) (255 mg, 0.99 mmol) was added to a solution of tert-butyl5-[[3-fluoro-4-[[2-(morpholine-4-carbothioylamino)acetyl]amino]phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(450 mg, 0.66 mmol) in CH₂Cl₂:THF (1:1, 20 mL). The reaction mixture wascooled to −30° C. and NH₃ gas purged for 15 minutes. The reactionmixture was stirred at RT for 4 h, quenched with MeOH (1 mL) andconcentrated under reduced pressure. Water (25 mL) was added andextracted with EtOAc (2×50 mL). The combined organic layer was driedover Na₂SO₄, filtered and concentrated. The crude material was purifiedby flash chromatography eluting with 3% MeOH in CH₂Cl₂ to afford a blackgummy material (250 mg, 57%).

M/z 663.53 (M+H)⁺

c.5-[[3-fluoro-4-[[2-(morpholine-4-carboximidoylamino)acetyl]amino]phenyl]sulfonylamino]thiazole-4-carboxylicacid

A solution of tert-butyl5-[[3-fluoro-4-[[2-(morpholine-4-carboximidoylamino)acetyl]amino]phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylatein TFA: H₂O (95:5, 2 mL) was stirred at RT for 3 h. The reaction mixturewas concentrated and the crude product was neutralised with methanolicammonia. Then the reaction mixture was concentrated under reducedpressure. The crude material was purified by preparative HPLC affordingthe title product as an off-white solid (12.4 mg, 8%).

¹H NMR (400 MHz, DMSO-d6) δ 13.32 (1H, brs), 9.67 (1H, s), 8.13 (1H, s),8.05-7.85 (3H, m), 7.63-7.61 (2H, m), 7.52-7.49 (1H, m), 4.18 (2H, brs),3.63-3.55 (4H, m), 3.50-3.30 (4H, obs).

M/z 487.34 (M+H)⁺

LC-MS Condition:

Column: Acquity BEH C18 (50 mm×2.1 mm, 1.7 um);

Mobile Phase: A: 0.05% Formic Acid in water; B: 0.05% Formic Acid inACN;

Time (min)/% B: 0/3, 0.4/3, 3.2/98, 3.8/98, 4.2/3, 4.5/3;

Column Temp: 35° C.,

Flow Rate: 0.6 mL/min.

Prep. HPLC Condition:

Column used: Symmetry C18 (300*19) mm, 7 u;

Mobile phase: (A) 0.1% Formic Acid (B) Acetonitrile;

Flow: 19 mL/min;

Gradient −(T/% B): 0/5, 1/5, 7.1/56, 7.15/99, 10/99, 10.1/5, 13/5;

Solubility: CH₃CN+H₂O.

Example 345-[[4-[[2-[(N-cyanocarbamimidoyl)amino]acetyl]amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylicacid

DIPEA (0.1 mL, 0.58 mmol) and NaN(CN)₂ (142 mg, 1.6 mmol) were added toa stirred solution of5-[[4-[(2-aminoacetyl)amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylicacid (200 mg, 0.53 mmol) in DMF (5 mL) at RT under nitrogen atmosphere.The reaction mixture was stirred at 50° C. for 48 h and concentratedunder reduced pressure. The crude compound was diluted with water (5 mL)and acidified to pH2-3 with 1N HCl. The resulting precipitate wasfiltered and dried under high vacuum. The crude product was purified bypreparative HPLC affording the title product as an off-white solid (20.8mg, 8%).

¹H NMR (400 MHz, DMSO-d₆) δ 12.8 (1H, brs), 10.0 (1H, s), 8.17 (1H, s),8.13-8.09 (1H, m), 7.55-7.52 (2H, m), 6.96 (1H, brs), 6.87 (2H, s), 4.00(2H, d, J=6.0 Hz).

M/z 442.18 (M+H)⁺

LC-MS Condition:

Column: Acquity BEH C18 (50 mm×2.1 mm, 1.7 um);

Mobile Phase: A: 0.05% Formic Acid in water; B: 0.05% Formic Acid inACN;

Time (min)/% B: 0/3, 0.4/3, 2/98, 3.4/98, 3.5/3, 4/3;

Column Temp: 35° C.,

Flow Rate: 0.6 mL/min.

Prep. HPLC Condition:

Column used: XBRIDGE C18 (150*19) mm, 5 u;

Mobile phase: (A) 0.1% Formic Acid (B) Acetonitrile;

Flow: 19 mL/min,

Gradient −(T/% B): 0/0, 2/0, 8/20, 10.9/20, 10.95/99, 13/99, 13.10/0,16/0;

Solubility: ACN+H₂O+THF.

Example 355-[[4-[(4-amino-4-imino-butanoyl)amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylicacid

a.5-[[4-(3-chloropropanoylamino)-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylicacid

Acetic anhydride (5 mL) in DCM (5 mL) was added to a stirred solution of3-chloropropanoyl chloride (1.5 g, 3.04 mmol) in DCM (5 mL) at 0° C.After 10 minutes, tert-butyl5-[(4-amino-3-fluoro-phenyl)sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(1.5 g) in DCM (10 mL) was added at 0° C. The resulting reaction mixturewas stirred at RT for 2 h, quenched with ice cooled water (20 mL) andextracted with DCM (2×10 mL). The organic layer was dried over Na2SO₄,filtered and concentrated. The crude material was purified by flashchromatography (eluting with 60% ethyl acetate in petroleum ether) toafford an off-white solid (600 mg, 33%).

M/z 584.40 (M+H)⁺

b. tert-butyl5-[[4-(3-cyanopropanoylamino)-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

Sodium cyanide (76 mg, 1.54 mmol) was added to a stirred solution of5-[[4-(3-chloropropanoylamino)-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylicacid (600 mg, 1.02 mmol) in DMF (5 mL) at RT. The resulting reactionmixture was stirred at RT for 6 h and quenched with ice cold water (10mL). The resulting precipitate was filtered, washed with Et₂O (2×10 mL)and dried under high vacuum to afford a brown solid (500 mg, 84%).

M/z 597.24 (M+Na)⁺

c. tert-butyl5-[[4-[(4-amino-4-imino-butanoyl)amino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

HCl gas was passed to a stirred solution of tert-butyl5-[[4-(3-cyanopropanoylamino)-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(400 mg, 0.69 mmol) in ethanol:Et₂O (1:4, 10 mL) at 0° C. for 2 h. Theresulting reaction mixture was kept at 4° C. for 16 h. Then the volatilecomponents were evaporated under reduced pressure. The residue wasdissolved in ethanol (5 mL) and NH₃ gas was passed for 20 minutes. Thevolatile components were evaporated under reduced pressure. Theresulting crude product was triturated with diethyl ether (2×5 mL) anddried under high vacuum to afford a brown solid which was used in thenext step without further purification (350 mg, crude).

M/z 592.43 (M+H)⁺

d.5-[[4-[(4-amino-4-imino-butanoyl)amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylicacid

TFA:H₂O (95:5, 3 mL) was added to tert-butyl5-[[4-[(4-amino-4-imino-butanoyl)amino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(350 mg, 0.592 mmol) at 0° C. The resulting reaction mixture was stirredat RT for 6 h and concentrated under reduced pressure. The resultingcrude product was triturated with diethyl ether (2×5 mL) and dried underhigh vacuum. The crude product was purified by preparative HPLCaffording the title product as an off-white solid (31 mg, 12%).

¹H NMR (400 MHz, DMSO-d₆) δ 13.40 (1H, s), 10.08 (1H, s), 8.99-8.71 (4H,m), 8.07 (1H, s), 8.05-8.01 (1H, m), 7.54-7.46 (2H, m), 2.85 (2H, t,J=7.2 Hz), 2.62 (2H, t, J=7.2 Hz).

M/z 415.93 (M+H)⁺

LC-MS Condition:

Column: Acquity BEH C18 (50 mm×2.1 mm, 1.7 um);

Mobile Phase: A: 0.05% Formic Acid in water; B: 0.05% Formic Acid inACN;

Time (min)/% B: 0/3, 0.4/3, 2/98, 3.4/98, 3.5/3, 4/3;

Column Temp: 35° C.;

Flow Rate: 0.6 mL/min.

Prep. HPLC Condition:

Column: Symmetry C18 (300*19) mm, 7 u;

Mobile phase: (A) 0.1% Formic Acid (B) Acetonitrile;

Flow: 19 mL/min;

Gradient (T/% B): 0/5, 1/5, 7/20, 10.1/20, 10.1/99, 13/99, 13.1/5, 16/5;

Solubility: ACN+H₂O+THF+DMSO+conc FA.

Example 365-[[4-[3-(4,5-dihydro-1H-imidazol-2-yl)propanoylamino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylicacid

a.5-[[4-[3-(4,5-dihydro-1H-imidazol-2-yl)propanoylamino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylicacid

HCl gas was passed to a stirred solution of tert-butyl5-[[4-(3-cyanopropanoylamino)-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(310 mg, 0.53 mmol) in ethanol:Et₂O (1:4, 15 mL) at 0° C. for 2 h. Theresulting reaction mixture was kept in refrigerator for 16 h. Then thevolatile components were evaporated under reduced pressure. Theresulting residue was dissolved in ethanol (5 mL). Then ethylene diamine(32 mg, 0.53 mmol) was added at RT. The resulting reaction mixture wasstirred at RT for 8 h and concentrated under reduced pressure. Theresulting crude product was triturated with n-pentane (2×5 mL) and driedunder high vacuum to afford a brown solid which was used in the nextstep without further purification (400 mg, crude).

M/z 618.46 (M+H)⁺

b.5-[[4-[3-(4,5-dihydro-1H-imidazol-2-yl)propanoylamino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylicacid

TFA:H₂O (95:5, 3 mL) was added to5-[[4-[3-(4,5-dihydro-1H-imidazol-2-yl)propanoylamino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylicacid (280 mg, 0.45 mmol) at 0° C. The resulting reaction mixture wasstirred at RT for 4 h and concentrated under reduced pressure. Theresulting crude product was triturated with diethyl ether (2×5 mL) anddried under high vacuum. The crude product was purified by preparativeHPLC to afford the title product as an off-white solid (21 mg, 10%).

¹H NMR (300 MHz, DMSO-d₆) δ 13.45 (1H, brs), 9.80 (1H, brs), 8.12-8.04(2H, m), 7.54-7.44 (2H, m), 3.65 (4H, s), 2.82-2.77 (2H, m), 2.62-2.58(2H, m).

M/z 441.98 (M+H)⁺

LC-MS Condition:

Column: Acquity BEH C18 (50 mm×2.1 mm, 1.7 um);

Mobile Phase: A: 0.05% Formic Acid in water; B: 0.05% Formic Acid inACN;

Time (min)/% B: 0/3, 0.4/3, 3.2/98, 3.8/98, 4.2/3, 4.5/3;

Column Temp: 35° C.;

Flow Rate: 0.6 mL/min.

Prep. HPLC Condition:

Column: Symmetry C18 (300*19) mm, 7 u;

Mobile phase: (A) 0.1% Formic Acid (B) Acetonitrile;

Flow: 19 mL/min;

Gradient (T/% B): 0/5, 1/5, 7/30, 8.7/30, 8.75/99, 11/99, 11.1/5, 13/5;

Solubility: ACN+H₂O+THF.

Example 375-[[4-[(3-amino-3-imino-2-methyl-propanoyl)amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylicacid

a. tert-butyl5-[[4-(2-chloropropanoylamino)-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

2-chloropropanoyl chloride (1.48 mL, 15.1 mmol) was added to a stirredsolution of tert-butyl5-[(4-amino-3-fluoro-phenyl)sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(3 g, 6.0 mmol) in DCM (50 mL) at 0° C. The resulting reaction mixturewas stirred at RT for 2 h and concentrated under reduced pressure. Theresidue was triturated with diethyl ether (2×50 mL), pentane (2×50 mL)and dried under reduced pressure to afford an off-white solid (3.4 g,95%).

M/z 606.28 (M+Na)⁺; 582.75 (M−H)⁻

b. tert-butyl5-[[4-(2-cyanopropanoylamino)-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

NaCN (570 mg, 11.6 mmol) was added to a solution of tert-butyl5-[[4-(2-chloropropanoylamino)-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(3.4 g, 5.8 mmol) in DMF (35 mL) at RT. The resulting reaction mixturewas stirred at RT for 16 h and quenched with ice cooled water. Theresulting precipitate was filtered and dried under high vacuum. Thecrude product was purified by silica gel chromatography (eluting with40% EtOAc in petroleum ether) to afford an off-white solid (1.5 g, 44%).

M/z 597.29 (M+Na)⁺; 573.62 (M−H)⁻

c. tert-butyl5-[[3-fluoro-4-[[3-(hydroxyamino)-3-imino-2-methyl-propanoyl]amino]phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

NH₂OH.HCl (362 mg, 5.22 mmol) and Na2CO3 (828 mg, 7.8 mmol) were addedto a stirred solution of tert-butyl5-[[4-(2-cyanopropanoylamino)-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(1.5 g, 2.61 mmol) in EtOH (30 mL) at RT. The resulting reaction mixturewas stirred at 65° C. for 1 h, cooled to RT and filtered. The filtratewas concentrated under reduced pressure to afford a pale yellow gummymaterial which was used in the next step without further purification(1.5 g, crude).

M/z 608.48 (M+H)⁺

d. tert-butyl5-[[4-[(3-amino-3-imino-2-methyl-propanoyl)amino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

Iron powder (193 mg, 3.4 mmol) was added to tert-butyl5-[[3-fluoro-4-[[3-(hydroxyamino)-3-imino-2-methyl-propanoyl]amino]phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(350 mg, 0.57 mmol) in ethanol:water (1:1, 3 mL) and heated to refluxfor 30 minutes. Then 1N HCl (0.3 mL) in ethanol:water (1:1, 3 mL) wasadded to the reaction mixture over a period of 30 minutes. The reactionmixture was stirred for an additional 1 h at 70° C., cooled to RT andfiltered through celite. The celite pad was washed with ethanol (2×10mL). The filtrate was concentrated under reduced pressure to afford apale yellow liquid which was used in the next step without furtherpurification (340 mg, crude).

M/z 592.28 (M+H)⁺

e.5-[[4-[(3-amino-3-imino-2-methyl-propanoyl)amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylicacid

Tert-butyl5-[[4-[(3-amino-3-imino-2-methyl-propanoyl)amino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(340 mg, 0.57 mmol) was added to a solution of TFA:H₂O (9:1, 3 mL) atRT. The reaction mixture was stirred at RT for 3 h and concentratedunder reduced pressure (below 30° C. of bath temperature). The residuewas triturated with diethyl ether (2×5 mL) and dried under high vacuum.The crude product was purified by preparative HPLC to afford the titleproduct as an off-white solid (48.2 mg).

¹H NMR (300 MHz, DMSO-d₆) δ 13.40 (1H, brs), 10.25 (1H, s), 8.89 (2H,s), 8.65 (2H, s), 8.11 (1H, s), 8.0 (1H, t, J=8.1 Hz), 7.60-7.50 (2H,m), 3.87 (1H, q, J=7.2 Hz), 1.49 (3H, d, J=7.2 Hz).

M/z 416.34 (M+H)⁺

LC-MS Condition:

Column: Acquity BEH C18 (50 mm×2.1 mm, 1.7 um);

Mobile Phase: A: 0.05% Formic Acid in water; B: 0.05% Formic Acid inACN;

Time (min)/% B: 0/3, 0.4/3, 3.2/98, 3.8/98, 4.2/3, 4.5/3;

Column Temp: 35° C.;

Flow Rate: 0.6 mL/min.

Prep. HPLC Condition:

Column used: Symmetry C18 (300*19) mm, 7 u;

Mobile phase: (A) 0.1% Formic Acid (B) Acetonitrile;

Flow: 19 mL/min;

Gradient −(T/% B): 0/5, 1/5, 8/50, 8.1/99, 11/99, 11.1/5, 14/5;

Solubility: ACN+H₂O+Concentrated FA.

Example 385-[[3-fluoro-4-[[2-(2-iminoimidazolidin-1-yl)acetyl]amino]phenyl]sulfonylamino]thiazole-4-carboxylicacid

a. tert-butyl5-[[3-fluoro-4-[[2-(2-thioxoimidazolidin-1-yl)acetyl]amino]phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

A solution of tert-butyl5-[[4-[(2-chloroacetyl)amino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(500 mg, 0.87 mmol) in acetonitrile (50 mL) was added to a stirredsolution of ethylenediamine (0.29 mL, 4.38 mmol) in acetonitrile (50 mL)over a period of 30 minutes at 75° C. The resulting reaction mixture wasstirred at 75° C. for 2.5 h. Di(imidazol-1-yl)methanethione (1.56 g,8.77 mmol) was then added at 75° C. and the reaction was stirred for 1 hat the same temperature then concentrated under reduced pressure. Theresulting crude compound was diluted with EtOAc (50 mL), washed withwater (10 mL) and brine solution (10 mL). The organic layer was driedover Na₂SO₄, filtered and concentrated under reduced pressure. The crudematerial was purified by flash chromatography (eluting with 60% EtOAc inpetroleum ether) to afford a light brown solid (200 mg, 36%).

M/z 636.20 (M+H)⁺

b. tert-butyl5-[[3-fluoro-4-[[2-(2-iminoimidazolidin-1-yl)acetyl]amino]phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

Ag(OTf) (121 mg, 0.47 mmol) and saturated NH₃ in THF (5 mL) were addedto a stirred solution of tert-butyl5-[[3-fluoro-4-[[2-(2-thioxoimidazolidin-1-yl)acetyl]amino]phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(200 mg, 0.31 mmol) in CH₂Cl₂ (10 mL) at −30° C. under nitrogenatmosphere. The resulting reaction mixture was stirred at RT for 16 h,quenched with MeOH (2 mL) and stirred at RT for 10 minutes. The reactionmixture was filtered through celite pad and the pad was washed withCH₂Cl₂ (2×10 mL). The filtrate was concentrated under reduced pressureto afford a dark brown liquid which was used in the next step withoutfurther purification (300 mg, crude).

M/z 619.48 (M+H)⁺

c.5-[[3-fluoro-4-[[2-(2-iminoimidazolidin-1-yl)acetyl]amino]phenyl]sulfonylamino]thiazole-4-carboxylicacid

TFA: H₂O (9:1, 5 mL) was added to tert-butyl5-[[3-fluoro-4-[[2-(2-iminoimidazolidin-1-yl)acetyl]amino]phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(270 mg, 0.43 mmol) at 0° C. The resulting reaction mixture was stirredat RT for 3 h and concentrated under reduced pressure. The resultingcrude product was triturated with diethyl ether (2×5 mL) and dried underhigh vacuum. The crude product was purified by preparative HPLCaffording the title product as an off-white solid (10.6 mg).

¹H NMR (400 MHz, DMSO-d₆) δ 13.32 (1H, brs), 10.24 (1H, s), 8.70 (1H,brs), 8.12 (1H, s), 7.68-7.60 (3H, m), 7.47 (1H, dd, J=8.0 Hz, J=7.6Hz), 7.38 (1H, s), 4.11 (2H, s), 3.74-3.67 (2H, m), 3.61-3.55 (2H, m).

M/z 443.24 (M+H)⁺

LC-MS Condition:

Column: Acquity BEH C18 (50 mm×2.1 mm, 1.7 um);

Mobile Phase: A: 0.05% Formic Acid in water; B: 0.05% Formic Acid inACN;

Time (min)/% B: 0/3, 0.4/3, 3.2/98, 3.8/98, 4.2/3, 4.5/3;

Column Temp: 35° C.,

Flow Rate: 0.6 mL/min

Prep. HPLC Condition:

Column used: Atlantis T3 (250*19) mm, 5 u;

Mobile phase: (A) 0.1% Formic Acid (B) Acetonitrile;

Flow: 19 mL/min;

Gradient −(T/% B): 0/5, 1/5, 7/30, 8.25/30, 8.3/99, 11/99, 11.1/5, 14/5;

Solubility: ACN+H₂O+THF

Example 395-[[4-[[2-[[N-(2-aminoethyl)carbamimidoyl]amino]acetyl]amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylicacid

a. tert-butyl5-[[4-[[2-[2-(tert-butoxycarbonylamino)ethylcarbamothioylamino]acetyl]amino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

Di(imidazol-1-yl)methanethione (97 mg, 0.54 mmol) was added to a stirredsolution of tert-butyl5-[[4-[(2-aminoacetyl)amino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(200 mg, 0.36 mmol) in DCM (10 mL) at RT. The reaction mixture wasstirred at RT for 4 h and NH₂CH₂CH₂NHBoc (174 mg, 1.08 mmol) was added.The reaction mixture was stirred at 40° C. for 6 h and concentratedunder reduced pressure. The crude product was purified by columnchromatography (eluting with 60% EtOAc in petroleum ether) to afford abrown solid (80 mg, 29%).

M/z 753.43 (M+H)⁺

b. tert-butyl5-[[4-[[2-[[N-[2-(tert-butoxycarbonylamino)ethyl]carbamimidoyl]amino]acetyl]amino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

Ag(OTf) (107 mg, 0.47 mmol) was added to a stirred solution oftert-butyl5-[[4-[[2-[2-(tert-butoxycarbonylamino)ethylcarbamothioylamino]acetyl]amino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(220 mg, 0.27 mmol) in CH₂Cl₂ (10 mL) at RT. The reaction mixture wasstirred at RT for 15 minutes and saturated NH₃ in THF (5 mL) was addedat −30° C. under nitrogen atmosphere. The resulting reaction mixture wasstirred at RT for 6 h, filtered through celite pad and the pad waswashed with CH₂Cl₂ (10 mL). The filtrate was concentrated and theobtained crude material was triturated with n-pentane (10 mL) to afforda brown solid which was used in the next step without furtherpurification.

M/z 736.40 (M+H)⁺

c.5-[[4-[[2-[[N-(2-aminoethyl)carbamimidoyl]amino]acetyl]amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylicacid

TFA: H₂O (9:1, 2 mL) was added to tert-butyl5-[[4-[[2-[[N-[2-(tert-butoxycarbonylamino)ethyl]carbamimidoyl]amino]acetyl]amino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(210 mg, 0.28 mmol) at 0° C. The reaction mixture was stirred at RT for4 h and concentrated under reduced pressure. The resulting crude productwas triturated with diethyl ether (2×5 mL) and dried under high vacuum.The crude product was purified by preparative HPLC affording the titleproduct as brown solid (25 mg).

¹H NMR (300 MHz, DMSO-d₆) δ 8.13 (1H, brs), 8.09 (1H, s), 7.42-7.30 (2H,m), 6.96 (1H, dd, J=8.7 Hz, J=8.4 Hz), 6.20-6.00 (1H, m), 5.70-5.40 (1H,m), 3.82 (2H, s), 3.27 (2H, brs), 2.80-2.75 (2H, m).

M/z 460.30 (M+H)⁺

LC-MS Condition:

Column: Acquity BEH C18 (50 mm×2.1 mm, 1.7 um);

Mobile Phase: A: 0.05% Formic Acid in water; B: 0.05% Formic Acid inACN;

Time (min)/% B: 0/3, 0.4/3, 3.2/98, 3.8/98, 4.2/3, 4.5/3;

Column Temp: 35° C.;

Flow Rate: 0.6 mL/min.

Prep. HPLC Condition:

Column: Atlantis T3 (250*19) mm, 5 u;

Mobile phase: (A) 0.1% Formic Acid (B) Acetonitrile;

Flow: 19 mL/min;

Gradient (T/% B): 0/5, 1/5, 7/25, 12/30, 12.1/99, 15/99, 15.1/5, 18/5;

Solubility: ACN+H₂O+THF+FA.

Compounds prepared using methods analogous to those described above forExample 39 by using methyl-amine in step-a and purified in a similarmanner by preparative HPLC are shown in the Table below:—

Example Structure Name, NMR and Mass 40

5-[[3-fluoro-4-[[2-[(N- methylcarbamimidoyl)amino]acetyl]amino]phenyl]sulfonylamino]thiazole-4-carboxylic acid M/z 431.2 (M + H)⁺ ¹HNMR (d6-DMSO) δ 13.39 (1H, s), 9.65 (1H, s), 8.11 (1H, m), 8.05 (1H, m),7.94 (3H, m), 7.61 (2H, m), 7.49 (1H, d, J = 8.4 Hz), 3.88 (2H, d, J =5.2 Hz), 2.65 (3H, d, J = 4.4 Hz).

Example 415-[[4-[[2-(2-carbamimidoylhydrazino)acetyl]amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylicacid

a. tert-butyl5-[[4-[[2-(2-tert-butoxycarbonylhydrazino)acetyl]amino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

KI (1.17 g, 7.01 mmol) was added to a solution of tert-butyl5-[[4-[(2-chloroacetyl)amino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(2 g, 3.50 mmol) in DMF (20 mL) at RT. After 10 minutes, tert-butylN-aminocarbamate (695 mg, 5.26 mmol) was added to the reaction mixtureat the same temperature. The resulting reaction mixture was stirred atRT for 16 h and concentrated under reduced pressure. Water (25 mL) wasadded to the crude compound and stirred for 20 minutes. The resultingprecipitate was filtered, washed with diethyl ether and dried under highvacuum to afford a pale yellow solid which was used in the next stepwithout further purification (1.2 g, 52%).

M/z 666.48 (M+H)⁺

b.5-[[3-fluoro-4-[(2-hydrazinoacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylicacid

TFA (4 mL) was added to tert-butyl5-[[4-[[2-(2-tert-butoxycarbonylhydrazino)acetyl]amino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(1.2 g, 1.80 mmol) at RT. The reaction mixture was stirred at RT for 16h and concentrated under reducing pressure. The resulting crude productwas triturated with diethyl ether (3×10 mL) to afford a pale yellowsolid which was used in the next step without further purification (1 g,crude).

M/z 390.32 (M+H)⁺

c.5-[[4-[[2-(2-carbamimidoylhydrazino)acetyl]amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylicacid

DIPEA (1.1 mL, 6.42 mmol) and pyrazole-1-carboxamidine; hydrochloride(212 mg, 1.92 mmol) were added to a stirred solution of5-[[3-fluoro-4-[(2-hydrazinoacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylicacid (500 mg, 1.28 mmol) in DMF (5 mL) at RT. The resulting reactionmixture was stirred at RT for 16 h, concentrated under reduced pressureand water (5 mL) was added to the residue. The resulting precipitate wasfiltered and washed with Et₂O (2×10 mL) and dried under high vacuum. Thecrude product was purified by preparative HPLC affording the titleproduct as an off-white solid (15 mg).

¹H NMR (400 MHz, DMSO-d₆) δ 13.40 (1H, brs), 10.0 (1H, brs), 9.00 (1H,brs), 8.48 (1H, s), 8.05 (1H, m), 7.55-7.49 (2H, m), 7.45-7.22 (3H,brs), 5.67 (1H, brs), 3.62 (2H, d, J=4.4 Hz).

M/z 432.37 (M+H)⁺

LC-MS Condition:

Column: Acquity BEH C18 (50 mm×2.1 mm, 1.7 um);

Mobile Phase: A: 0.05% Formic Acid in water; B: 0.05% Formic Acid inACN;

Time (min)/% B: 0/3, 0.4/3, 3.2/98, 3.8/98, 4.2/3, 4.5/3;

Column Temp: 35° C.;

Flow Rate: 0.6 mL/min.

Prep. HPLC Condition:

Column: X BRIDGE C18 (150*19) mm, 5 u;

Mobile phase (A) 0.1% Formic Acid (B) Acetonitrile;

Flow: 19 mL/min;

Gradient (T/% B): (T/% B): 0/0, 3/0, 8.8/33, 9/33, 9.10/99, 12/99,12.10/0, 15/0;

Solubility: ACN+H₂O+THF+DMSO+FA.

Example 425-[[3-fluoro-4-[(2-guanidinooxyacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylicacid

a. tert-butyl5-[[4-[[2-(tert-butoxycarbonylamino)oxyacetyl]amino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

A solution of tert-butyl N-hydroxycarbamate (175 mg, 1.31 mmol) in THF(10 mL) was added to NaH (195 mg, 4.38 mmol) suspension in THF (10 mL)at 0° C. under argon atmosphere and stirred at RT for 30 minutes. Thentert-butyl5-[[4-[(2-chloroacetyl)amino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(500 mg, 0.87 mmol) in THF (10 mL) was added to above reaction mixtureat 0° C. under argon atmosphere. The resulting reaction mixture wasstirred at RT for 1.5 h, quenched with ice cold water (20 mL) andextracted with ethyl acetate (2×20 mL). The combined organic layer wasdried over Na₂SO₄, filtered and concentrated under reduced pressure. Thecrude material was purified by column chromatography (eluting with 30%ethyl acetate in petroleum ether) to afford a yellow solid (350 mg,59%).

M/z 667.10 (M+H)⁺

b.5-[[4-[(2-aminooxyacetyl)amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylicacid

TFA: H₂O (9:1, 3 mL) was added to tert-butyl5-[[4-[[2-(tert-butoxycarbonylamino)oxyacetyl]amino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(300 mg, 0.44 mmol) at 0° C. The reaction mixture was stirred at RT for4 h and concentrated under reduced pressure. The resulting crude productwas triturated with diethyl ether (3×10 mL) to afford an off-white solidwhich was used in next step without further purification (200 mg,crude).

M/z 390.95 (M+H)⁺

c.5-[[3-fluoro-4-[(2-guanidinooxyacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylicacid

DIPEA (0.26 mL, 1.53 mmol) and pyrazole-1-carboxamidine; hydrochloride(149 mg, 0.92 mmol) were added to a stirred solution of5-[[4-[(2-aminooxyacetyl)amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylicacid (200 mg, 0.51 mmol) in DMF (6 mL) at 0° C. The resulting reactionmixture was stirred at RT for 6 h, concentrated under reduced pressureand water (5 mL) was added to the residue. The resulting precipitate wasfiltered, washed with Et₂O (2×10 mL) and dried under high vacuum. Thecrude product was purified by preparative HPLC affording the titleproduct as an off-white solid (70 mg, 31%).

¹H NMR (300 MHz, DMSO-d₆) δ 13.50 (1H, brs), δ 9.60 (1H, s), 8.37 (2H,brs), 8.17-8.12 (1H, m), 8.07 (1H, s), 7.58-7.51 (2H, m), 5.45 (2H,brs), 4.62 (2H, brs), 4.22 (2H, s).

M/z 433.30 (M+H)⁺

LC-MS Condition:

Column: Acquity BEH C18 (50 mm×2.1 mm, 1.7 um);

Mobile Phase: A: 0.05% Formic Acid in Water B: 0.05% Formic Acid inAcetonitrile;

Time (min)/% B: 0/3, 0.4/3, 3.2/98, 3.8/98, 4.2/3, 4.5/3;

Column Temp: 35° C.;

Flow Rate: 0.6 mL/min.

Prep. HPLC Condition:

Column: X BRIDGE C18 (150*19) mm, 5 u;

Mobile phase (A) 0.1% Formic Acid (B) Acetonitrile;

Flow: 19 mL/min;

Gradient (T/% B): −(T/% B):0/0, 3/0, 8.8/33, 9/33, 9.10/99, 12/99,12.10/0, 15/0;

Solubility: ACN+H₂O+THF+DMSO+FA.

Example 435-[[4-[[(2E)-2-(carbamimidoylhydrazono)acetyl]amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylicacid

a. tert-butyl5-[[4-[[4-[4-[(4-tert-butoxycarbonylthiazol-5-yl)-[(4-methoxyphenyl)methyl]sulfamoyl]-2-fluoro-anilino]-4-oxo-but-2-enoyl]amino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

But-2-enedioyl dichloride (0.18 g, 1.2 mmol) in DCM (20 mL) was added toa solution of tert-butyl5-[(4-amino-3-fluoro-phenyl)sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(1 g, 2.0 mmol) in DCM (10 mL) at 0° C. The resulting reaction mixturewas stirred at RT for 6 h, diluted with DCM and washed with water (2×10mL) and brine (2×10 mL) solution. The combined organic layer was driedover Na₂SO₄, filtered and concentrated under reduced pressure. The crudematerial was purified by flash chromatography (eluting with 50% ethylacetate in petroleum ether) to afford a pale yellow solid (500 mg, 23%).

M/z 1067.05 (M+H)⁺

b. tert-butyl5-[[3-fluoro-4-(oxaldehydoylamino)phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

A solution of tert-butyl5-[[4-[[4-[4-[(4-tert-butoxycarbonylthiazol-5-yl)-[(4-methoxyphenyl)methyl]sulfamoyl]-2-fluoro-anilino]-4-oxo-but-2-enoyl]amino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(500 mg, 0.46 mmol) in DCM/MeOH (3:1, 40 mL) was purged with ozone gasfor 1 h at −78° C. Then DMS (2 mL) was added to the reaction mixture atthe same temperature and the resulting mixture was stirred at RT for 2h. The crude reaction mixture was used in next step without furtherpurification.

c. tert-butyl5-[[4-[[(2E)-2-(carbamimidoylhydrazono)acetyl]amino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

1-aminoguanidine; hydrochloride (30 mg, 0.27 mmol) was added to astirred solution of tert-butyl5-[[3-fluoro-4-(oxaldehydoylamino)phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(100 mg, 0.18 mmol) in DCM/MeOH (1:1, 10 mL) at RT. The resultingreaction mixture was stirred at RT for 16 h and concentrated underreduced pressure. The crude product was used in next step withoutfurther purification (100 mg, crude).

M/z 522.1 (M+H)⁺

d.5-[[4-[[(2E)-2-(carbamimidoylhydrazono)acetyl]amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylicacid

TFA/H₂O (9:1, 1.0 mL) was added to tert-butyl5-[[4-[[(2E)-2-(carbamimidoylhydrazono)acetyl]amino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(80 mg, 0.13 mmol) at 0° C., stirred at RT for 4 h and concentratedunder reduced pressure. The resulting crude product was triturated withdiethyl ether (2×5 mL) and dried under high vacuum. The crude productwas purified by preparative HPLC affording the title product as anoff-white solid (11 mg).

¹H NMR (400 MHz, DMSO-d₆) δ 9.68 (1H, brs), 8.05 (1H, s), 7.80 (1H, dd,J=8.4 Hz, J=8.0 Hz), 7.53-7.47 (2H, m), 7.19 (1H, s), 6.80-6.00 (4H,brs).

M/z 430.31 (M+H)⁺

LC-MS Condition:

Column: Acquity BEH C18 (50 mm×2.1 mm, 1.7 um);

Mobile Phase: A: 0.05% Formic Acid in water; B: 0.05% Formic Acid inACN;

Time (min)/% B: 0/3, 0.4/3, 3.2/98, 3.8/98, 4.2/3, 4.5/3;

Column Temp: 35° C.;

Flow Rate: 0.6 mL/min.

Prep. HPLC Condition:

Column: Atlantis T3 (250*19) mm, 5 u;

Mobile phase: (A) 0.1% Formic Acid (B) Acetonitrile;

Flow: 19 mL/min;

Gradient (T/% B): 0/5, 1/5, 7/25, 12/30, 12.1/99, 15/99, 15.1/5, 18/5;

Solubility: ACN+H₂O+THF+FA

Example 44 5-[(4-guanidinophenyl)sulfonylamino]thiazole-4-carboxylicacid

a. tert-butyl5-[[4-[[N,N′-bis(tert-butoxycarbonyl)carbamimidoyl]amino]phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

A solution of tert-butyl5-[(4-aminophenyl)sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(500 mg, 1.05 mmol) in THF (20 mL) was added to NaH (250 mg, 10.5 mmol)suspension in THF (20 mL) at 0° C. under argon atmosphere. After 30minutes, a solution of tert-butylN-[(tert-butoxycarbonylamino)-pyrazol-1-yl-methylene]carbamate (1.0 g,3.43 mmol) in THF (10 mL) was added at 0° C. under argon atmosphere. Theresulting reaction mixture was stirred at RT for 16 h, quenched with icecold water (20 mL) and extracted with ethyl acetate (2×20 mL). Thecombined organic layer was dried over Na₂SO₄, filtered and concentratedunder reduced pressure. The crude material was purified by triturationwith diethyl ether (2×5 mL) to afford a pale yellow solid which was usedin the next step without further purification (150 mg, crude).

M/z 662.03 (M+H-Boc)⁺

b. 5-[(4-guanidinophenyl)sulfonylamino]thiazole-4-carboxylic acid

TFA:H₂O (9:1, 2 mL) was added to tert-butyl5-[[4-[[N,N′-bis(tert-butoxycarbonyl)carbamimidoyl]amino]phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(150 mg, 0.22 mmol) at RT. The resulting mixture was stirred for 4 h andconcentrated under reduced pressure. The resulting crude product wastriturated with diethyl ether (2×5 mL) and dried under high vacuum. Thecrude product was purified by preparative HPLC affording the titleproduct as an off-white solid (22 mg, 28%).

¹H NMR (400 MHz, DMSO-d₆) δ 13.60 (1H, brs), 8.05 (1H, s), 7.74 (2H, d,J=8.8 Hz), 7.47 (3H, brs), 7.25 (2H, d, J=8.8 Hz).

M/z 342.29 (M+H)⁺

LC-MS Condition:

Column: Acquity BEH C18 (50 mm×2.1 mm, 1.7 um);

Mobile Phase: A: 0.05% Formic Acid in water; B: 0.05% Formic Acid inACN;

Time (min)/% B: 0/3, 0.4/3, 3.2/98, 3.8/98, 4.2/3, 4.5/3;

Column Temp: 35° C.,

Flow Rate: 0.6 mL/min.

Prep. HPLC Condition:

Column: Symmetry C18 (300*19) mm, 7 u;

Mobile phase: (A) 0.1% Formic Acid (B) Acetonitrile;

Flow: 19 mL/min;

Gradient (T/% B): 0/2, 1/2, 8/30, 9.10/99, 12/99, 12.10/2, 15/2;

Solubility: ACN+H₂O+DMSO.

Example 455-[[3-fluoro-4-[[(2-guanidinoacetyl)amino]methyl]phenyl]sulfonylamino]thiazole-4-carboxylicacid

a. tert-butyl5-[(3-fluoro-4-vinyl-phenyl)sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

A solution of tert-butyl5-[(4-bromo-3-fluoro-phenyl)sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(3 g, 5.38 mmol) in 1,4-dioxane (40 mL) was purged with argon for 15minutes. Then 4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborolane (0.99 g,6.45 mmol), K₂CO3 (1.11 g, 8.07 mmol), PdCl₂(PPh₃)₂ (0.37 g, 0.53 mmol)were added under argon atmosphere. The resulting reaction mixture washeated to 85° C. for 24 h in a closed vial. The reaction mixturetemperature was allowed to cool to RT, filtered through a celite pad(washed with EtOAc (2×50 mL)). The organic layer was concentrated underreduced pressure. The resulting crude compound was dissolved in ethylacetate (50 mL), washed with water (50 mL) and brine solution (50 mL).The organic layer was dried over Na₂SO₄, filtered and concentrated undervacuum. The crude compound was purified by flash chromatography (elutingwith 20% ethyl acetate in petroleum ether) affording an off-white solid(1.5 g, 55%).

M/z 505.1 (M+H)⁺

b. tert-butyl5-[(3-fluoro-4-formyl-phenyl)sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

NaIO₄ (8.51 g, 39.8 mmol) and OSO₄ (1.68 g, 6.63 mmol) were added to asolution of tert-butyl5-[(3-fluoro-4-vinyl-phenyl)sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(6.7 g, 13.2 mmol) in acetonitrile:H₂O:CCl4 (1:1:1, 60 mL). Theresulting reaction mixture was stirred at RT for 4 h. Water (30 mL) wasadded and the mixture was extracted with ethyl acetate (2×100 mL). Thecombined organic extracts were dried over Na₂SO₄, filtered andconcentrated. The crude material was purified by flash chromatography(eluting with 22% ethyl acetate in petroleum ether) affording anoff-white solid (4.5 g, 66%).

M/z 507.4 (M+H)⁺

c. tert-butyl5-[[3-fluoro-4-[hydroxyiminomethyl]phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

Tert-butyl5-[(3-fluoro-4-formyl-phenyl)sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(3.6 g, 7.10 mmol) solution in EtOH (20 mL) was added to a stirredsolution of hydroxylamine hydrochloride (593.8 mg, 8.52 mmol) andammonium chloride (454.9 mg, 8.52 mmol) in H₂O:EtOH (4:1, 30 mL). Theresulting reaction mixture was stirred at RT for 4 h. Water (20 mL) wasadded and the mixture extracted with ethyl acetate (2×50 mL). Thecombined organic extracts were dried over Na₂SO₄, filtered andconcentrated. The crude material was purified by flash chromatography(eluting with 40% ethyl acetate in petroleum ether) affording anoff-white solid (2.8 g, 75%).

M/z 522.1 (M+H)⁺

d. tert-butyl5-[[4-(aminomethyl)-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

Zn dust (0.52 g, 8.04 mmol) was added to a stirred solution oftert-butyl5-[[3-fluoro-4-[hydroxyiminomethyl]phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(2.8 g, 5.36 mmol) in AcOH (20 mL) at RT. The resulting reaction mixturewas stirred at RT for 16 h. Ice cold water was added and the mixture wasextracted with ethyl acetate (2×50 mL). The combined organic extractswere dried over Na₂SO₄, filtered and concentrated. The crude materialwas purified by trituration with diethyl ether (2×10 mL) affording ayellow solid (2 g, 73%).

M/z 508.1 (M+H)⁺

e. tert-butyl5-[[4-[[[2-(tert-butoxycarbonylamino)acetyl]amino]methyl]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

DIPEA (0.61 mL, 3.54 mmol) and HATU (0.67 g, 1.77 mmol) were added to asolution of tert-butyl5-[[4-(aminomethyl)-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(0.6 g, 1.18 mmol) and 2-(tert-butoxycarbonylamino)acetic acid (0.31 g,1.77 mmol) in DMF (20 mL) under argon atmosphere. The resulting reactionmixture was stirred at RT for 4 h. Ice cold water (10 mL) was added andthe mixture was extracted with ethyl acetate (2×20 mL). The combinedorganic extracts were dried over Na₂SO₄, filtered and concentrated. Thecrude material was purified by flash chromatography (eluting with 50%ethyl acetate in petroleum ether) affording an off-white solid (500 mg,63%).

M/z 508.1 (M+H)⁺

f.5-[[4-[[(2-aminoacetyl)amino]methyl]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylicacid, trifluoroacetate

TFA (5 mL) was added to tert-butyl5-[[4-[[[2-(tert-butoxycarbonylamino)acetyl]amino]methyl]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate(500 mg, 0.75 mmol) at RT and stirred for 4 h. TFA was evaporated byreduced pressure. The resulting crude product was triturated withdiethyl ether (2×10 mL) and dried under high vacuum affording anoff-white solid (250 mg, 85%).

M/z 389.1 (M+H)⁺

g.5-[[3-fluoro-4-[[(2-guanidinoacetyl)amino]methyl]phenyl]sulfonylamino]thiazole-4-carboxylicacid

Pyrazole-1-carboxamidine, hydrochloride (141.2 mg, 0.96 mmol) and DIPEA(0.55 mL, 3.21 mmol) were added to a stirred solution of5-[[4-[[(2-aminoacetyl)amino]methyl]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylicacid, trifluoroacetate (250 mg, 0.64 mmol) in DMF (6 mL) at RT. Theresulting reaction mixture was stirred at RT for 18 h and concentratedunder reduced pressure. Water (5 mL) was added to the residue. Theresulting precipitate was filtered and washed with diethyl ether (2×5mL). The crude product was purified by preparative HPLC to afford thetitle compound as a white solid (70 mg, 25%).

¹H NMR (500 MHz, DMSO-d₆) δ 13.43 (1H, brs), 8.60 (1H, brs), 8.08 (1H,s), 7.51-7.42 (3H, m), 7.50-7.10 (4H, brs), 4.33 (2H, s), 3.85 (2H, s).

M/z 431.0 (M+H)⁺

LC-MS Condition:

Column: Acquity UPLC BEH C18 (50 mm×2.1 mm, 1.7 um);

Mobile Phase: A: 0.1% Formic Acid in Water; B: 0.1% Formic Acid inAcetonitrile;

Flow Rate: 0.8 mL/min

Time (min)/% B: 0/2, 0.4/2, 2.2/98, 2.6/98, 2.61/2, 3.0/2.

Column Temp: 60° C.

Prep. HPLC Condition:

Column: KROMASIL-C18 (150*25 MM), 10 u;

Mobile phase: 0.05% Formic acid in H₂O: ACETONITRILE;

Flow: 25 mL/min

Gradient (T/% B): 0/5, 1/5, 7/40, 7.1/98, 9/98, 9.1/5, 11/5;

Solubility: ACN+H₂O+THF+DMSO

Compounds prepared using methods analogous to those described above forExample 45 and purified in a similar manner by preparative HPLC areshown in the Table below:—

Example Structure Name, NMR and Mass 46

5-[[3-fluoro-4-(guanidinomethyl) phenyl]sulfonylamino]thiazole-4-carboxylic acid M/z 374.0 (M + H)⁺ ¹H NMR (d6-DMSO) δ 13.40 (1H, s),8.09 (1H, s), 7.59 (1H, d, J = 7.5 Hz), 7.47 (1H, d, J = 10 Hz), 7.59(1H, d, J = 7.5 Hz), 7.50-7.25 (4H, brs), 4.41 (2H, s).

Example 47: Activity of Compounds of the Invention

Experiments were conducted to determine:

-   -   (1) The inhibitory activity of the compounds of the invention        against MBL enzymes;    -   (2) The plasma protein binding for compounds of the invention;        and    -   (3) The plasma stability of compounds of the invention.

Details of the protocols used for each of the sets of experiments areset out below:

1. Enzymatic Inhibition

In Vitro Enzyme Inhibition Assays

Enzyme inhibition assays were performed using purified MBL enzymes(NDM-1; VIM-1; VIM-2; IMP-1) in 10 mM HEPES buffer pH 7.5 in 96-wellmicrotiter plates. Imipenem (300 μM) was used as substrate and itshydrolysis was followed at UV 299 nm during 10 mn every 30 seconds usinga Perkin Elmer Envision UV fluorescence plate reader. Hydrolysis ratedata in presence of a range of inhibitors was analysed using Dotmaticsdatabase software and calculated IC₅₀ values were converted to Ki valuesusing the Cheng-Prusoff equation:Ki=IC ₅₀/(1+([S]/K _(m))where the K_(m) values for NDM-1, VIM-2 and IMP-1 are 70 μM, 1.5 μM, 9μM and 25 μM respectively. Compound dilution was performed in DMSO.

Mean Ki values from multiple experiments are presented below.Experimental results are shown given using the following bands:

-   -   For NDM-1 the Ki values of <0.05 μM are designated (A); Ki        values of 0.05-0.2 μM are designated (B); Ki values of >0.2 μM        (0.2-2 μM) are designated (C).    -   For VIM-1 the Ki values of <0.2 μM are designated (A); Ki values        of 0.2-0.5 μM are designated (B); Ki values of >0.5 μM (0.5-1        μM) are designated (C).    -   For VIM-2 the Ki values of <0.02 μM are designated (A); Ki        values of 0.02-0.05 μM are designated (B); Ki values of >0.05 μM        (0.05-0.15 μM) are designated (C).    -   For IMP-1 the Ki values of <0.5 μM are designated (A); Ki values        of 0.5-1 μM are designated (B); Ki values of >1 μM (1-10 μM) are        designated (C).

Ki Values for Compounds of the Invention.

Ki/μM Example NDM-1 VIM-1 VIM-2 IMP-1 2 (A) (B) (C) (C) 3 (B) (B) (B)(C) 4 (A) (B) (B) (C) 5 (B) (A) (B) (A) 6 (C) (B) (C) 7 (B) (B) (C) (C)8 (B) (B) (C) 9 (B) (B) (A) (C) 10 (A) (A) (A) (B) 11 (C) (B) (C) 12 (B)(B) (B) 13 (A) (A) (C) (B) 14 (C) (B) (C) 15 (B) (C) (C) 16 (B) (A) (A)(B) 17 (B) (A) (A) (A) 18 (B) (A) (A) (A) 19 (B) (B) (B) (C) 20 (B) (B)21 (A) (B) (B) 22 (A) (B) (C) (C) 23 (A) (A) (B) (C) 24 (B) (C) (C) 25(B) (B) (B) 26 (A) (B) (B) (C) 27 (A) (A) (A) (B) 28 (B) (A) (A) (A) 29(A) (B) (B) (C) 30 (A) (B) (B) (C) 31 (A) (A) (B) (A) 32 (B) (B) (B) (C)33 (C) (B) (C) 34 (A) (C) (B) (A) 35 (A) (A) (B) (B) 36 (A) (B) (C) (C)37 (C) (A) (C) 38 (C) (B) (C) 39 (C) (A) (B) (C) 40 (C) (B) (C) 41 (A)(B) (B) (B) 42 (C) (C) (C) 43 (B) (B) (A) 44 (C) (B) (C) (C) 45 (C) (C)(C) (C) 46 (B) (B) (C) (C)

2. Antimicrobial Susceptibility Testing

Antibiotic Activity of β-Lactam Antibiotics on MBL Expressing Bacteriain the Presence of the Compounds of the Invention

The experiments were carried out using the ‘broth micro-dilution method’according to the protocols M07-A8 established by the Clinical LaboratoryStandards Institute (CLSI). Serial dilutions of the β-lactam antibiotic(Meropenem) were prepared in 96-well plates in cation-adjustedMueller-Hinton broth (CAMHB); the concentration range was defined from0.03 mg/L to 512 mg/L. The compounds were added at a constantconcentration of 8 μg/mL. A bacterial inoculum of each strain (clinicalisolates) was adjusted to a 0.5 McFarland turbidity standard inphysiologic serum (0.9% NaCl), then diluted 1:100 in CAMHB and added toeach well to give a final bacterial cell number of 5×10⁵ CFU/well. Afterincubation for 18-20 hours in a heating chamber at 37° C., the growthinhibition was evaluated by the absence of any bacterial development.

Minimal inhibitory concentrations (MIC) are taken as the lowestconcentration of antibiotic at which the test organism did not showvisible growth; results were confirmed by measuring the optical density(OD) at 600 nm in a spectrophotometer.

Compounds of the invention were tested at a constant concentration of 8μg/mL. The clinical strains used in these potentiation experiments wereNTBC020 (E. coli strain expressing NDM-1, TEM-1 and CTX-M-15); NTBC035-2(K. pneumoniae strain expressing NDM-1, CMY-4 and SHV-11); NTBC104-1 (K.pneumoniae strain expressing NDM-1 and SHV-11); NTBC123 (K. pneumoniaestrain expressing NDM-1); NTBC018 (C. freundii strain expressing VIM-2);NTBC024 (K. pneumoniae strain expressing VIM-19, TEM-1 and CTX-M-3);NTBC042 (E. coli strain expressing VIM-1, TEM-1, CTX-M-15, SHV-12);NTBC055 (E. Coli strain expressing VIM-1); NTBC062 (K. pneumoniae strainexpressing IMP-1 and TEM-1) and NTBC039 (K. oxytoca strain expressingIMP-28).

Results are shown below. Data are banded as follows: MIC values of <1μg/mL are designated (A); MIC values of 1-2 μg/mL are designated (B);MIC values of >2 g/mL (2-200 μg/mL) are designated (C).

Strain Example NTBC020 NTBC035-2 NTBC104-1 NTBC123 NTBC018 NTBC024NTBC042 NTBC055 NTBC062 NTBC039 2 (B) (B) (C) (C) (A) (B) (A) (B) (B)(B) 3 (C) (B) (B) (C) (A) (B) (A) (B) (B) (C) 4 (B) (B) (C) (C) (A) (B)(A) (B) (B) (B) 5 (B) (B) (C) (C) (A) (A) (A) (B) (B) (A) 7 (B) (B) (B)(C) (A) (B) (A) (B) (B) (B) 10 (C) (B) (C) (C) (A) (A) (A) (B) (B) (B)13 (B) (B) (C) (C) (A) (B) (A) (B) (B) (B) 15 (C) (C) (C) (C) (B) (C)(C) (C) (C) (C) 16 (C) (C) (C) (C) (A) (A) (A) (A) (B) (B) 17 (C) (C)(C) (C) (A) (A) (A) (B) (B) (A) 19 (B) (B) (C) (C) (A) (B) (A) (B) (B)(A) 20 (C) (C) (C) (C) (A) (C) (A) (B) (B) (B) 22 (B) (B) (C) (C) (A)(C) (B) (C) (B) (B) 23 (C) (B) (A) (C) (A) (B) (B) (B) (B) (C) 24 (C)(C) (C) (C) (A) (B) (A) (B) (C) (C) 25 (C) (C) (C) (C) (A) (C) (B) (B)(B) (B) 26 (A) (A) (B) (C) (A) (A) (A) (B) (B) (C) 27 (C) (C) (C) (C)(B) (B) (A) (B) (B) (B) 29 (C) (B) (C) (C) (A) (B) (A) (C) (B) (B) 30(C) (C) (B) (C) (A) (A) (A) (B) (B) (B) 32 (B) (B) (C) (C) (A) (B) (A)(B) (B) (B) 34 (C) (C) (C) (C) (A) (B) (A) (B) (B) (B) 35 (C) (B) (B)(C) (A) (B) (A) (B) (B) (B) 36 (C) (C) (C) (C) (A) (C) (A) (B) (B) (B)38 (C) (C) (C) (C) (B) (C) (B) (C) (B) (C) 41 (B) (B) (C) (C) (A) (B)(A) (B) (B) (B) 43 (C) (C) (C) (C) (A) (B) (B) (B) (B) (C) 44 (C) (C)(C) (C) (A) (B) (A) (A) (B) (C) 45 (C) (C) (C) (C) (B) (C) (B) (B) (B)(B) 46 (C) (B) (C) (C) (B) (B) (A) (A) (B) (C)

Example 48: Comparative Study

3. Plasma Protein Binding

Protocol Summary

Method Rapid equilibrium dialysis Species Human Plasma Plasma 100%plasma Test compound concentration 10 μM Buffer Phosphate buffer salinepH 7.4 Incubation Time 5 h No of replicates 2 QC Compounds WarfarinFinal DMSO Concentration <0.1% Analytical Method LC-MS/MS

Assay Procedure

Test compound was spiked in plasma to a final concentration of 10 μM. Analiquot of 300 μL of plasma was placed in red chamber of the insert and500 μL of PBS was placed into white chamber of the insert. The plate wasincubated at 37° C. in thermomixer at 400 rpm for 5 hours. Afterincubation, the samples were matrix equilibrated with opposite matrix(10 μL of plasma/100 μL of buffer sample was matched with 100 μL ofblank buffer/10 μL of plasma). Matrix matched samples were precipitatedwith 200 μL of acetonitrile containing internal standard. Samples werevortexed at 1000 rpm for 5 min and centrifuged at 4000 rpm for 10 min.Supernatant was separated, diluted 2 fold with water and analyzed onLC-MS/MS. Blank control samples were processed immediately after thepreparation of plasma working stock solutions. These samples served as ameasure for calculating the % recovery of test compounds.

Data Analysis and Calculation

The percent plasma bound fraction was calculated by the followingequations:% Unbound=100*F _(C) /T _(C)% Recovery=100*(F _(C) +T _(C))/T ₀where

-   T_(C)=Total compound concentration as determined by the calculated    concentration on the plasma side of the membrane-   F_(C)=Free compound concentration as determined by the calculated    concentration on the buffer side of the membrane-   T₀=Total compound concentration as determined before dialysis

For each set of duplicates/compound, the percentage bound, percentageunbound and percentage recovery was determined. Results are as shownbelow.

4. Plasma Stability of Test Compounds

Protocol Summary

Test compound concentration 1 μM Matrix Human Plasma Incubation Time 0,1, 3 and 5 h No of replicates 2 per each time point QC CompoundsPropantheline Analytical Method LC-MS/MS End point % Remaining of thetest compound

Assay Procedure

Test compound and QC compound were incubated at a final concentration of1 μM in plasma at 37° C. in shaker water bath with gentle shaking. Atpredetermined time points, reaction was terminated with 200 μL ofacetonitrile containing internal standard and centrifuged at 4000×RCF,4° C. for 20 minutes. Supernatant was separated and analyzed byLC-MS/MS.

Data Analysis and Calculation

The following equation was used to determine the percentage remaining oftest/QC compound following the procedure above:

$\%\mspace{14mu}{remaining}\mspace{14mu}{of}\mspace{14mu}{the}\mspace{14mu}{test}\mspace{14mu}{substance}{= {\frac{{Peak}\mspace{14mu}{Area}\mspace{14mu}{ratio}\mspace{14mu}{at}\mspace{14mu}{time}\mspace{11mu}\left( \min \right)}{{Peak}\mspace{14mu}{Area}\mspace{14mu}{Ratio}\mspace{14mu}{at}\mspace{14mu} 0\mspace{14mu}\min} \times 100}}$

Results are as shown below.

Studies were undertaken to compare the compounds of the invention withstructurally similar compounds (“Comp. x”). Experiments were conductedas described above. Data were banded as set out below.

-   -   For VIM-1 the Ki values of <0.15 μM are designated (++++); Ki        values of 0.15-0.3 μM are designated (+++); Ki values of 0.3-0.5        μM are designated (++); Ki values of >0.5 μM (0.5-1 μM) are        designated (+).    -   For IMP-1 the Ki values of <0.15 μM are designated (++++); Ki        values of 0.15-0.6 μM are designated (+++); Ki values of 0.6-5        μM are designated (++); Ki values of >5 μM (5-10 μM) are        designated (+).    -   For VIM-2 the Ki values of <0.02 μM are designated (++++); Ki        values of 0.02-0.05 μM are designated (+++); Ki values of        0.05-0.1 μM are designated (++); Ki values of >0.1 μM (0.1-0.15        μM) are designated (+).    -   For NDM-1 the Ki values of <0.03 μM are designated (++++); Ki        values of 0.03-0.1 μM are designated (+++); Ki values of 0.1-0.3        are designated (++); Ki values of >0.5 μM (0.3-2 μM) are        designated (+).

Human Stability in plasma human protein Example plasma binding Comp. A

29% remaining after 2 hours 90.3% Example 7

100% remaining after 5 hours 60.8% Comp. B

0% remaining after 5 hours Not available Example 2

73% remaining after 2 hours 81% Comp. E

29% remaining after 2 hours Not available Example 16

89% remaining after 2 hours Not available

MBL-inhibitory efficacy was observed for the compound of Example 7.

Ki/μM VIM- IMP- VIM- NDM- Example 1 1 2 1 Comp. B

++ ++ + ++++ Example 2

++ ++ ++ ++++ Comp. C

+ ++ + ++ Example 3

++ ++ +++ ++ Comp. D

+ +++ +++ +++ Example 5

+++ +++ +++ +++ Comp. E

++ ++ ND ++ Example 16

++++ +++ ++++ +++ Comp. F

+++ ND +++ +++ Example 17

++++ ++++ ++++ +++ Comp. G

++ +++ +++ +++ Example 18

++++ ++++ ++++ +++ Comp. H

+ + +++ ++ Example 20

+++ ND ND ++ Comp. I

+ ++ ND ++ Example 29

+++ ++ +++ ++++ ND: Not determined

The compounds of the invention were also found to exhibit better enzymeinhibition (lower Ki values) and better potentiation (lower MIC values)against the above mentioned MBL enzymes (VIM/IMP/NDM) and strains ofbacteria as compared to the structurally related analogues lacking a—C(NR)—NR₂ motif.

As can be readily seen, the compounds of the invention are associatedwith improved properties compared with structurally analogous compounds.This finding is surprising, not least because the —C(NR)—NR₂ motifcommon to the compounds of the invention can be associated with rapidhydrolysis, so may have been expected to render the compounds unsuitablefor the type of use described herein. The fact that this motif can beintroduced not only without prejudicing the efficacy of the compounds,but also with an enhancement of plasma stability and efficacy, is thusunexpected.

5. PK (Pharmacokinetic) Studies.

Compound A and Example 7 were dosed i.v. at 1 mg/kg to male Swiss albinomice. The measured PK parameters are shown in the Table below:—

Compound C₀ (ng/mL) AUC (ng · h/mL) Cl (mL/min/kg) Compound A  17   9819 Example 7 1814 1036  16 C₀ = plasma concentration AUC = area underthe curve Cl = clearance

Note that for the same dosage:

-   -   1. Example 7 achieves over a 100-fold maximum concentration        compared to Compound A;    -   2. Example 7 achieves over a 100-fold exposure (AUC, integration        of concentration vs time) compared to Compound A; and    -   3. Example 7 is cleared from the blood about 50-fold slower than        Compound A.

This data is in accord with the in vitro data generated on plasmastability and confirms plasma stability as the limiting factor withregard to the potential of Compound A to be useful in animal efficacystudies.

6. In Vivo Efficacy Studies

Mice were infected in the thigh with K. pneumoniae NTBC104. The MIC ofmeropenem against this strain is 64 ug/mL due to the strain producingNDM-1. The MIC of meropenem in the presence of 8 ug/mL of the compoundof Example 2 is 4 ug/mL.

At the end of the experiment (9 hours post infection) the animals weresacrificed and the numbers of colony forming units (CFUs) were measuredin order to quantify bacterial load (extent of the infection). Meropenemat 30 mg/kg reduced bacterial load slightly whereas meropenem at 30mg/kg plus the compound of Example 2 at 30 mg/kg significantly reducedthe bacterial load compared to meropenem alone, showing a 1.6 Log₁₀reduction in CFUs. Results are shown in FIG. 1.

Under the same experimental conditions the compound of Example 7effected a 1.7 Log₁₀ reduction in CFUs compared to meropenem alone.Compound A was not progressed to efficacy studies as the in vitro and PKstudies predicted this compound would fail in efficacy studies so it isnot ethical to carry out such an experiment.

Under the same conditions the compound of Example 26 effected a 1.8Log₁₀ reduction in CFUs compared to meropenem alone.

7. Extended MIC Profiling Vs MBL-Expressing Clinical Strains

To assess the coverage and potentiation of meropenem by compounds of theinvention, the susceptibility of around 200 clinical isolates wasexamined. The criteria for selection into the panel was that theclinical strain was resistant to carbapenems, but only expressing NDMenzyme variants and not serine betalactamase enzymes with carbapenemaseactivity such as KPC or OXA.

At 8 μg/mL concentration of either Example 2 or Example 26, meropenem ispotentiated to the extent that just under 90% of strains exhibit ameropenem MIC of 8 ug/mL, whereas the same concentration of meropenemalone is only stopping the growth of <1% of the strains and within theparameters of this experiment the cessation of growth of 90% of allstrains could not be achieved with meropenem alone. Results are shown inFIG. 2.

Example 49

1. Combination Therapy by Compounds of the Invention with SBL Inhibitorsand Antibiotic Agents

As discussed above, bacteria exhibit resistance to antibiotics bymechanisms including both the modification of the biological target suchthat binding affinity for the antibiotic is reduced, and the productionof enzymes which deactivate the antibacterial drug, such asbeta-lactamase enzymes (including both serine-β-lactamases, SBL, andmetallo-β-lactamases, MBL). A proposed strategy to address suchresistance is to administer combination therapies comprising agentswhich inhibit the enzymes which deactivate the antibiotic together withthe antibiotic itself. In other words, it may be possible to rescue theantibacterial activity of the drug by using a dual combination approachof antibiotic plus a drug that inhibits the deactivating enzyme

Combinations of serine β-lactamase inhibitors with antibiotics areknown. For example, the Streptomyces natural product clavulanic acid, aserine β-lactamase inhibitor, was developed as a dual combinationtogether with the β-lactam antibiotic amoxicillin under the nameAugmentin. More recently avibactam, a serine β-lactamase inhibitor withan improved spectrum of serine β-lactamase inhibition over clavulanicacid, has been introduced into the clinic in combination with thecephalosporin β-lactam antibiotic ceftazidime (known together asAvycaz). However, these combinations are ineffective at treatingbacterial infection caused by bacteria which express MBL enzymes, as theSBL inhibitors are typically inactive against such enzymes.

A further complication of having two distinct categories of β-lactamaseenzymes present in bacterial infections arises as neither diagnostictests to very rapidly ascertain the precise mechanism of β-lactamaseresistance nor dual inhibitors of both serine and metallo β-lactamaseenzymes are currently available in the clinic. Indeed, at present noclinically-approved metallo β-lactamase inhibitor to address the problemof metallo β-lactamase enzymes exists, even if a rapid diagnostic testwas available to allow resistance due to SBL enzymes to be distinguishedfrom that due to MBL enzymes.

The inventors have now recognised that a product which is apharmaceutical combination of an antibiotic, serine β-lactamaseinhibitor and a metallo β-lactamase inhibitor (a so-called triplecombination) could overcome the need to identify if a resistantbacterium causing a particular infection was producing a serineβ-lactamase or a metallo β-lactamase enzyme (or both, in an increasingnumber of very resistant strains). In this regard, there are threepossible scenarios for the β-lactamase profile of carbapenem-resistantenterobacteriaceae (CRE). Group 1 organisms have either exclusivelymetallo β-lactamase enzymes or a mixture of metallo β-lactamase andserine β-lactamase enzymes but the resistance is primarily due to themetallo β-lactamase. Group 2 organisms have serine β-lactamase enzymesonly. Group 3 organisms have both metallo β-lactamase and serineβ-lactamase enzymes and both enzymes play a significant role inresistance.

The following abbreviations are used in this Example:—

CMY: Class C β-lactamase

TEM: Class A β-lactamase

SHV: Class A B-lactamase (sulfhydryl variable)

CTX-M: Class A β-lactamase (CTX for cefotaximase and M for Munich)

OSBL: “older-spectrum” β-lactamases

OXA: Class D β-lactamase (oxacillinase)

ACT-TYPE: Class C β-lactamase (AmpC-type beta-lactamase)

KPC: Class A β-lactamase (K. pneumoniae carbapenemase)

VIM: Verona integron-encoded metallo-β-lactamase

NDM: New Delhi metallo-β-lactamase

IMP: Imipenemase metallo-β-lactamase

Experiments were carried out using the ‘broth micro-dilution method’according to the protocols M07-A8 established by the Clinical LaboratoryStandards Institute (CLSI). Serial dilutions of meropenem (mero) wereprepared in 96-well plates in cation-adjusted Mueller-Hinton broth(CAMHB); the concentration range was defined from 0.03 mg/L to 512 mg/L.The compounds (the compound of Example 2, above, and/or WCK4234) wereadded at the concentration indicated in the table below. A bacterialinoculum of each strain (clinical isolates) was adjusted to a 0.5McFarland turbidity standard in physiologic serum (0.9% NaCl), thendiluted 1:100 in CAMHB and added to each well to give a final bacterialcell number of 5×10⁵ CFU/well. After incubation for 18-20 hours in aheating chamber at 37° C., the growth inhibition was evaluated by theabsence of any bacterial development.

Minimum inhibitory concentrations (MIC) are taken as the lowestconcentration of antibiotic at which the test organism did not showvisible growth; results were confirmed by measuring the optical density(OD) at 600 nm in a spectrophotometer.

The SBL inhibitor WCK4234 was synthesized according to the proceduredescribed in WO 2015/114595.

In brief, WCK4234 and its sodium salt were synthesised followingpublished procedures (WO2105114595), the latter stages of which areshown below:—

The compound of Formula (B) was prepared by the synthesis described indetail by Ball, M. et al in Organic Process Research and Development,(2016), 1799.

The clinical strains used in these experiments were as follows:

Group 1 (Strains where the Resistance is Primarily Due to Metalloβ-Lactamase Enzymes):

NTBC020 (E. coli strain expressing NDM-1, TEM-1 and CTX-M-15); NTBC035-2(K. pneumoniae strain expressing NDM-1, CMY-4 and SHV-11); NTBC104-1 (K.pneumoniae strain expressing NDM-1 and SHV-11); NTBC123 (K. pneumoniaestrain expressing NDM-1); NTBC062 (K. pneumoniae strain expressing IMP-1and TEM-1); NTBC024 (K. pneumoniae strain expressing VIM-19, TEM-1 andCTX-M-3); NTBC042 (E. coli strain expressing VIM-1, TEM-1, CTX-M-15,SHV-12); NTBC055 (E. Coli strain expressing VIM-1); and NTBC039 (K.oxytoca strain expressing IMP-28).

Group 2 (Strains where the Resistance is Due to Serine β-LactamaseEnzymes):

NTBC091-1 (E. coli strain expressing KPC-2 and TEM-1); NTBC093 (E.cloacae strain expressing KPC-2 and TEM-1); NTBC096-1 (K. pneumoniastrain expressing OXA-181 and SHV-11); NTBC099 (K. pneumonia strainexpressing KPC-3, SHV-11 and TEM-1); and NTBC189 (K. pneumonia strainexpressing TEM-OSBL, CTX-M-14 and OXA-48).

Group 3 (Strains where the Resistance is Due to Both Serine and Metalloβ-Lactamase Enzymes):

NTBC019 (K. pneumonia strain expressing NDM-1, CTX-M-15 and OXA-181);NTBC185 (K. pneumonia strain expressing SHV-OSBL, TEM-OSBL, NDM-1 andOXA-48); NTBC186 (K. pneumonia strain expressing ACT-TYPE, VIM-1 andOXA-48); NTBC187 (K. pneumonia strain expressing SHV-OSBL, NDM-1 andOXA-48); and NTBC188 (K. pneumonia strain expressing NDM-1 and KPC-2).

Results are shown below. Data are banded as follows: MIC values of <1μg/mL are designated (A); MIC values of 1 or 2 g/mL are designated (B);MIC values of 4 or 8 μg/mL are designated (C); and MIC values ≥16 μg/mLare designated (D).

MIC mero + mero + WCK4234 mero/ WCK4234 mero + (4 ug/mL) + Strain μg/mL(4 μg/mL) Ex. 2 (8 μg/mL) Ex. 2 (8 μg/mL) Group 1 NTBC020 128 (D) (A)(A) NTBC035-2 64 (D) (B) (B) NTBC104-1 64 (D) (A) (A) NTBC123 128 (D)(C) (C) NTBC062 4 (C) (B) (B) NTBC024 16 (D) (A) (A) NTBC042 8 (C) (B)(B) NTBC055 4 (C) (B) (B) Group 2 NTBC091-1 4 (A) NTBC093 128 (A)NTBC096-1 16 (A) NTBC099 128 (A) NTBC189 16 (A) Group 3 NTBC019 64 (D)(B) (A) NTBC185 128 (D) (D) (C) NTBC186 16 (C) (C) (C) NTBC187 128 (D)(D) (C) NTBC188 32 (C) (C) (A)

As can be seen for Group 1 and Group 2 strains, the dual combination ofmeropenem and appropriate β-lactamase inhibitor reduces the MICrequired.

For Group 3 organisms the results indicate that the combination of a MBLinhibitor according to the invention and a SBL inhibitor (WCK4234)together with meropenem was capable of reducing the MIC required.

Discussion

In the antibacterial field there is no known example of triple therapyspecifically for the eradication of bacterial infection. There is oneknown example of triple therapy for the management of gastro-oesophagealreflux disease (GORD) where H. pylori infection is suspected to be acomponent in the disorder as well as gastric ulcers, but in this casethe National Institute of Clinical Excellence (NICE) guidelinesrecommend treatment with a triple combination of an anti-ulcer protonpump inhibitor with two antibiotics (amoxicillin and clarithromycin).Neither in development nor in the clinic are there any triplecombination of antibacterial drugs or antibacterial drugs plus adjuvantssuch as a β-lactamase enzyme inhibitor.

One significant advantage offered by the triple combination of theinvention is that when a CRE strain is encountered and rapid treatmentis essential for the survival of the patient, the use of the triplecombination means that in principle it is not essential to wait formicrobiological and molecular characterisation of the resistanceelements before commencing treatment. Thus, the triple combinationdescribed herein is useful in the prevention or treatment of anybacterial infection since it avoids the need for prior identification ofthe bacterial strain.

2. Further Data

Additional experiments were performed to demonstrate the advantages ofthe triple combination of the invention.

Experiments were conducted as described above. Compounds (the compoundsof Examples 2 and 26) were tested at 8 μg/mL. Avibactam and Wck4234 weretested at 4 μg/mL. MIC values were determined for

Strains tested expressed both carbapenemase one from class B (MBL) andone from class A or D (serine beta lactamase).

NTBC19 is K. pneumoniae expressing NDM-1; CTXM-15 and OXA-181.

NTBC188 is an E. cloacae expressing NDM-1 and KPC-2.

Data are banded as follows: MIC values of <0.5 μg/mL are designated (A);MIC values of 1-4 μg/mL are designated (B); MIC values of >8 μg/mL(8-512 μg/mL) are designated (C). Results are shown in the Table below.

NTBC19 NTBC188 Meropenem (C) (C) Meropenem + Avibactam (C) (C)Meropenem + Wck4234 (C) (C) Meropenem + Example 2 (B) (C) Meropenem +Example 2 + Avibactam (A) (A) Meropenem + Example 2 + WCK4234 (A) (A)Meropenem + Example 26 (B) (C) Meropenem + Example 26 + Avibactam (A)(A) Meropenem + Example 26 + WCK4234 (A) (A)

The data clearly show that the triple combination of (i) meropenem; (ii)a compound of the invention such as the compound of Example 2 or thecompound of Example 26; and (iii) an SBL inhibitor such as avibactam orWCK4234 beneficially leads to decreased MIC values in both strainstested.

The invention claimed is:
 1. A compound of Formula (I), or apharmaceutically acceptable salt thereof,

wherein R¹ is selected from H, R^(1a) and —CH₂OC(O)R^(1a), whereinR^(1a) is selected from an unsubstituted C₁ to C₄ alkyl group andphenyl; {circle around (A)} is a cyclic group selected from C₆ to C₁₀aryl and 5- to 10-membered heteroaryl; each R² is independently selectedfrom: (i) halo or R⁸; (ii) C₁₋₃ alkyl, O(C₁₋₃ alkyl), S(C₁₋₃ alkyl),SO(C₁₋₃ alkyl) or SO₂(C₁₋₃ alkyl), any of which may optionally besubstituted with 1, 2 or 3 halo substituents and/or one R′substituent;and (iii) NR^(a)C(O)R^(c), and NR^(a)C(O)NR^(b)R^(c), wherein each R^(a)and R^(b) is independently selected from hydrogen and unsubstituted C₁₋₂alkyl and each R^(c) is unsubstituted C₁₋₂ alkyl; and each R⁸ isindependently selected from CN, OH, —C(O)NR^(f)R^(g), —NR^(f)R^(g),—NR¹⁰C(NR¹¹)R¹², —C(NR¹⁰)NR¹¹R¹², and —NR¹⁰C(NR¹¹)NR¹²R¹³; wherein eachof R^(f) and R^(g) is independently H or unsubstituted C₁₋₂ alkyl; m is0, 1, 2 or 3 R³ is selected from hydrogen and a C₁ to C₃ alkyl groupwhich is unsubstituted or is substituted with 1, 2 or 3 substituentsselected from halogen, —OR¹⁰, and —NR¹⁰R¹¹; n is 0 or 1 Z is a bond oris selected from —NR¹⁰C(O)—, —C(O)NR¹⁰—, —NR¹⁰C(O)NR¹¹—, —NR¹⁰C(O)O—,—OC(O)NR¹⁰, —NR¹⁰C(O)S—, —SC(O)NR¹⁰, —NR¹⁰C(NR¹¹)—, —C(NR¹⁰)NR¹¹—,—NR¹⁰C(NR¹¹)NR¹²—, —NR¹⁰C(N⁺R¹¹R¹²)—, —C(N⁺R¹⁰R¹¹)NR¹²—,—NR¹⁰C(N⁺R¹¹R¹²)NR¹³—, —NR¹⁰C(NR¹¹)O—, —OC(NR¹⁰)NR¹¹,—NR¹⁰C(N⁺R¹¹R¹²)O—, —OC(N⁺R¹⁰R¹¹)NR¹²—, —NR¹⁰C(NR¹¹)S—, —SC(NR¹⁰)NR¹¹,—NR¹⁰C(N⁺R¹¹R¹²)S—, —SC(N⁺R¹⁰R¹¹)NR¹²—, —C(O)NR¹⁵—, —NR¹⁰C(O)NR¹⁵—,—OC(O)NR¹⁵, —SC(O)NR¹⁵, —C(NR¹⁰)NR¹⁵—, —NR¹⁰C(NR¹¹)NR¹⁵—,—C(N⁺R¹⁰R¹¹)NR¹⁵—, —NR¹⁰C(N⁺R¹¹R¹²)NR¹⁵—, —OC(NR¹⁰)NR¹⁵,—OC(N⁺R¹⁰R¹¹)NR¹⁵—, —SC(NR¹⁰)NR¹⁵, and —SC(N⁺R¹⁰R¹¹)NR¹⁵—; L is a bondor is selected from C₁₋₄ alkylene, C₂₋₄ alkenylene, C₂₋₄ alkynylene,C₁₋₃ alkylene-(C₃₋₆cycloalkylene)-C₁₋₃ alkylene, C₁₋₄alkylene-(C₃₋₆cycloalkylene) and (C₃₋₆cycloalkylene)-C₁₋₄ alkylene,wherein L is unsubstituted or is substituted with 1 or 2 substituentsselected from halogen, —OR¹⁰, and —NR¹⁰R₁₁; or L is —C(R¹⁰)═N—; X is abond or, when L is other than a bond or —C(R¹⁰)═N—, X is a bond or isselected from —NR¹⁰—, —O—, —NR¹⁰C(NR¹¹)—, and —C(NR¹⁰)—; p is 0 or 1; R⁴is selected from H, —CN and C₁ to C₃ alkyl which is unsubstituted or issubstituted with 1, 2 or 3 substituents selected from halogen, —OR¹⁰,—NR¹⁰R¹¹, and —CN; R⁵ is selected from H, —CN and C₁ to C₃ alkyl whichis unsubstituted or is substituted with 1, 2 or 3 substituents selectedfrom halogen, —OR¹⁰, —NR¹⁰R¹¹, and —CN; R⁶ is selected from H, —CN andC₁ to C₃ alkyl which is unsubstituted or is substituted with 1, 2 or 3substituents selected from halogen, —OR¹⁰, —NR¹⁰R¹¹, and —CN; R⁷ ifpresent is selected from H, —CN and C₁ to C₃ alkyl which isunsubstituted or is substituted with 1, 2 or 3 substituents selectedfrom halogen, —OR¹⁰, —NR¹⁰R¹¹, and —CN; each R¹⁰, R¹¹, R¹², and R¹³ isindependently H or methyl; each R¹⁵ is independently substituted C₁ toC₄ alkyl or unsubstituted C₂ to C₄ alkyl, wherein when R¹⁵ is asubstituted C₁ to C₄ alkyl group, the alkyl group is substituted with 1,2 or 3 substituents independently selected from halogen, CN, OR¹⁰ and—NR¹⁰R¹¹.
 2. The compound according to claim 1 wherein R¹ is H.
 3. Thecompound according to claim 1, wherein {circle around (A)} is a cyclicgroup selected from phenyl and 5- to 6-membered heteroaryl.
 4. Thecompound according to claim 1 wherein {circle around (A)} is selectedfrom phenyl, pyridazine, pyridine and thiazole.
 5. The compoundaccording to claim 1 wherein each R² is independently selected from: (i)halo, CN, OH, —C(O)NR^(f)R^(g), or —NR^(f)R^(g); wherein each of R^(f)and R^(g) is independently H or methyl; and (ii) C₁₋₂ alkyl, O(C₁₋₂alkyl), S(C₁₋₂ alkyl), or SO(C₁₋₂ alkyl); any of which may optionally besubstituted with 1, 2 or 3 halo substituents and/or one substituentselected from CN and OH.
 6. The compound according to claim 1 wherein R³is H.
 7. The compound according to claim 1 wherein n is
 0. 8. Thecompound according to claim 1 wherein Z is a bond or is selected from—NR¹⁰C(O)—, —C(O)NR¹⁰—, —NR¹⁰C(O)NR¹¹—, —NR¹⁰C(O)O—, —OC(O)NR¹⁰,—NR¹⁰C(O)S—, —SC(O)NR¹⁰, —NR¹⁰C(NR¹¹)—, —C(NR¹⁰)NR¹¹—, and—NR¹⁰C(NR¹¹)NR¹²—.
 9. The compound according to claim 1 wherein Z isselected from —NR¹⁰C(O)—, —C(O)NR¹⁰—, and —NR¹⁰C(O)NR¹¹—.
 10. Thecompound according to claim 1 wherein L is a bond or is selected fromC₁₋₄ alkylene, C₂₋₄ alkenylene, and C₂₋₄ alkynylene; or L is —C(R¹⁰)═N—.11. The compound according to claim 1 wherein L is selected from C₁₋₃alkylene and C₂₋₃ alkenylene.
 12. The compound according to claim 1wherein X is a bond.
 13. The compound according to claim 1 wherein p is1 and R⁷ is H or methyl.
 14. The compound according to claim 1 whereinR⁴ is H.
 15. The compound according to claim 1 wherein R⁵ is selectedfrom H, —CN and C₁ to C₂ alkyl which is unsubstituted or is substitutedwith 1, 2 or 3 halo substituents and/or one —NR¹⁰R¹¹ substituent and R⁶is H or methyl.
 16. The compound according to claim 1 wherein: R¹ is H;{circle around (A)} is a cyclic group selected from phenyl and 5- to6-membered heteroaryl; m is 0, 1 or 2; each R² is independently selectedfrom: halo or R⁸; C₁₋₂ alkyl, O(C₁₋₂ alkyl), S(C₁₋₂ alkyl), SO(C₁₋₂alkyl) or SO₂(C₁₋₂ alkyl), any of which may optionally be substitutedwith 1, 2 or 3 halo substituents and/or one R⁸ substituent; andNR^(a)C(O)R^(c), and NR^(a)C(O)NR^(b)R^(c), wherein each R^(a) and R^(b)is independently selected from hydrogen and unsubstituted C₁₋₂ alkyl andeach R^(c) is unsubstituted C₁₋₂ alkyl; each R⁸ is independentlyselected from CN, OH, —C(O)NR^(f)R^(g), and —NR^(f)R^(g); wherein eachof R^(f) and R^(g) is independently H or unsubstituted C₁₋₂ alkyl; n is0; or n is 1 and R³ is H Z is selected from —NR¹⁰C(O)—, —C(O)NR¹⁰—,—NR¹⁰C(O)NR¹¹—, —NR¹⁰C(O)O—, —OC(O)NR¹⁰, —NR¹⁰C(O)S—, —SC(O)NR¹⁰,—NR¹⁰C(NR¹¹)—, —C(NR¹⁰)NR¹¹—, and —NR¹⁰C(NR¹¹)NR¹²—; L is a bond or isselected from C₁₋₄ alkylene, C₂₋₄ alkenylene and C₂₋₄ alkynylene; or Lis —C(R¹⁰)═N—; X is a bond; i) p is 0; R⁴ is H and R⁵ is selected fromH, —CN and C₁ to C₂ alkyl which is unsubstituted or is substituted with1, 2 or 3 halo substituents and/or one —NR¹⁰R¹¹ substituent; and R⁶ is Hor methyl; or ii) p is 1; and R⁴ is H; R⁵ is selected from H, —CN and C₁to C₂ alkyl which is unsubstituted or is substituted with 1, 2 or 3 halosubstituents and/or one —NR¹⁰R¹¹ substituent; R⁶ is H or methyl and R⁷is H or methyl.
 17. The compound according to claim 1 wherein: R¹ is H;{circle around (A)} is selected from phenyl, pyridazine, pyridine andthiazole; m is 1 or 2; each R² is independently selected from: or halo,CN, OH, —C(O)NR^(f)R^(g), or —NR^(f)R^(g); wherein each of R^(f) and R⁹is independently H or methyl; and C₁₋₂ alkyl, O(C₁₋₂ alkyl), S(C₁₋₂alkyl), or SO(C₁₋₂ alkyl); any of which may optionally be substitutedwith 1, 2 or 3 substituents selected from halo, CN, OH; n is 0; Z isselected from —NR¹⁰C(O)—, —C(O)NR¹⁰—, and —NR¹⁰C(O)NR¹¹—; L is selectedfrom C₁₋₃ alkylene and C₂₋₃ alkenylene; X is a bond; p is 0; or p is 1and R⁷ is H; R⁴ is H; R⁵ is selected from H, —CN and C₁ to C₂ alkylwhich is unsubstituted or is substituted with 1, 2 or 3 halosubstituents and/or one —NR¹⁰R¹¹ substituent; and R⁶ is H.
 18. Thecompound according to claim 1 wherein R⁴, R⁵, R⁶ and R⁷ if present areeach hydrogen.
 19. The compound according to claim 1, wherein: R¹ isselected from H, R^(1a) and —CH₂OC(O)R^(1a), wherein R^(1a) is selectedfrom an unsubstituted C₁ to C₄ alkyl group and phenyl; {circle around(A)} is phenyl; m is 2; each R² is independently a halo group; n is 0; Zis —NR¹⁰C(O)NR¹¹—; L is a bond; X is a bond; p is 1; R⁷ is H; R⁴ is H;R⁵ is H; R⁶ is H; and R¹⁰ and R¹¹ are each H.
 20. The compound accordingto claim 1, which compound is selected from:5-[[4-[(2-guanidinoacetyl)amino]-3-(trifluoromethoxy)phenyl]sulfonylamino]thiazole-4-carboxylic acid;5-[[3-fluoro-4-[[(2-guanidinoacetyl)amino]methyl]phenyl]sulfonylamino]thiazole-4-carboxylicacid;5-[[3-fluoro-4-(guanidinomethyl)phenyl]sulfonylamino]thiazole-4-carboxylicacid;5-[[3-fluoro-4-(2-guanidinoethylsulfanylcarbonylamino)phenyl]sulfonylamino]thiazole-4-carboxylic acid;5-[[4-[2-[(2-amino-2-imino-ethyl)amino]-2-oxo-ethyl]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylicacid;5-[[3-carbamoyl-4-[(2-guanidinoacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylicacid;5-[[3-cyano-4-[(2-guanidinoacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylicacid;5-[[3-fluoro-4-(2-guanidinoethoxycarbonylamino)phenyl]sulfonylamino]thiazole-4-carboxylicacid; 5-[(4-guanidinophenyl)sulfonylamino]thiazole-4-carboxylic acid;5-[[4-[2-(2-carbamimidoylhydrazino)-2-oxo-ethyl]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylicacid;5-[[3-chloro-4-[(2-guanidinoacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylicacid;5-[[4-[(2-guanidinoacetyl)amino]-3-methoxy-phenyl]sulfonylamino]thiazole-4-carboxylicacid;5-[[4-[[2-(2-carbamimidoylhydrazino)acetyl]amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylicacid;5-[[4-[[(2E)-2-(carbamimidoylhydrazono)acetyl]amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylicacid;5-[[4-[[2-(4,5-dihydro-1H-imidazol-2-ylamino)acetyl]amino]-3,5-difluoro-phenyl]sulfonylamino]thiazole-4-carboxylicacid;5-[[6-[(2-guanidinoacetyl)amino]pyridazin-3-yl]sulfonylamino]thiazole-4-carboxylicacid;5-[[4-[(2-amino-2-imino-ethyl)carbamoylamino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylicacid;5-[[4-[(3-amino-3-imino-propanoyl)amino]-3,5-difluoro-phenyl]sulfonylamino]thiazole-4-carboxylic acid;5-[[4-[[3-(dimethylamino)-3-imino-propanoyl]amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylicacid;5-[[3-fluoro-4-[(2-guanidinooxyacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylicacid;5-[[3-fluoro-4-[[3-imino-3-(methylamino)propanoyl]amino]phenyl]sulfonylamino]thiazole-4-carboxylicacid;5-[[4-[3-(4,5-dihydro-1H-imidazol-2-yl)propanoylamino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylicacid;5-[[2-[(2-guanidinoacetyl)amino]thiazol-5-yl]sulfonylamino]thiazole-4-carboxylicacid;5-[[4-[[2-[(N-cyanocarbamimidoyl)amino]acetyl]amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylicacid;5-[[3-fluoro-4-(guanidinocarbamoylamino)phenyl]sulfonylamino]thiazole-4-carboxylicacid;5-[[3-fluoro-4-[[2-(morpholine-4-carboximidoylamino)acetyl]amino]phenyl]sulfonylamino]thiazole-4-carboxylicacid;5-[[4-[(3-amino-3-imino-2-methyl-propanoyl)amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylicacid;5-[[4-[[2-(4,5-dihydro-1H-imidazol-2-yl)acetyl]amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylicacid;5-[[4-(carbamimidoylcarbamoylamino)-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylicacid;5-[[4-[[(2R)-2-guanidinopropanoyl]amino]phenyl]sulfonylamino]thiazole-4-carboxylicacid;5-[[3,5-difluoro-4-[(2-guanidinoacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylicacid;5-[[4-[(4-amino-4-imino-butanoyl)amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylicacid;5-[[4-[[2-(4,5-dihydro-1H-imidazol-2-ylamino)acetyl]amino]-2,5-difluoro-phenyl]sulfonylamino]thiazole-4-carboxylicacid;5-[[2,5-difluoro-4-[(2-guanidinoacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylicacid;5-[[3-fluoro-4-[[2-[(N-methylcarbamimidoyl)amino]acetyl]amino]phenyl]sulfonylamino]thiazole-4-carboxylicacid;5-[[3-fluoro-4-[[2-(2-iminoimidazobdin-1-yl)acetyl]amino]phenyl]sulfonylamino]thiazole-4-carboxylicacid;5-[[4-[[2-[carbamimidoyl(methyl)amino]acetyl]amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylicacid;5-[[4-[[2-[[N-(2-aminoethyl)carbamimidoyl]amino]acetyl]amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylicacid;5-[[5-fluoro-6-[(2-guanidinoacetyl)amino]-3-pyridyl]sulfonylamino]thiazole-4-carboxylicacid;5-[[3-fluoro-4-(3-guanidinopropanoylamino)phenyl]sulfonylamino]thiazole-4-carboxylicacid;5-[[4-[(3-amino-3-imino-propanoyl)amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylicacid;5-[[3-fluoro-4-[(2-guanidinoacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylicacid; and5-[[4-[(2-guanidinoacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylicacid; and pharmaceutically acceptable salts thereof.
 21. Apharmaceutical composition comprising the compound according to claim 1together with at least one pharmaceutically acceptable carrier ordiluent.
 22. The pharmaceutical composition according to claim 21further comprising an antibiotic agent.
 23. The pharmaceuticalcomposition according to claim 22 wherein the antibiotic agent is aβ-lactam antibiotic.
 24. The pharmaceutical composition according toclaim 23 wherein the β-lactam antibiotic is selected from carbapenems,penicillins, cephalosporins and penems.
 25. The pharmaceuticalcomposition according to claim 23 wherein the β-lactam antibiotic ismeropenem.
 26. The pharmaceutical composition according to claim 21further comprising a serine-β-lactamase inhibitor.
 27. Thepharmaceutical composition according to claim 26 wherein theserine-β-lactamase inhibitor is a compound of Formula (II) or apharmaceutically acceptable salt thereof,

wherein G is selected from —CN and —C(O)NR^(j)R^(k); R^(k) is selectedfrom —W and -Q-W; wherein W is selected from 5- to 6-memberedheterocyclyl, R^(j) and —N(R^(j))₂; and Q is selected from —NR^(j)C(O)—,—C(O)—NR^(j)—, C₁₋₃ alkylene, —O—C₁₋₃ alkylene and —N(R^(j))—C₁₋₃alkylene; each R^(j) is selected from H and unsubstituted C₁₋₃ alkyl.28. The pharmaceutical composition according to claim 26 wherein theserine-β-lactamase inhibitor is selected from WCK4234, avibactam,relebactam, zidebactam and nacubactam, or pharmaceutically acceptablesalts thereof.